KR20210132645A - Method for preparing poly(alkyl cyanoacrylate)-based nano/microfiber and use thereof - Google Patents

Abstract

The present invention relates to the field of biomaterials, and more particularly to the field of poly(alkyl cyanoacrylate) based nano- and microfibers. The present invention provides a novel process for the preparation of ready-to-use poly(alkyl cyanoacrylate)-based nano/microfibers, which process is produced by anionic polymerization of alkyl cyanoacrylate monomers or oligomers and has a specific polydispersity. electrospinning of poly(alkyl cyanoacrylate) homopolymers or copolymers characterized by a degree index. Accordingly, the present invention also provides novel ready-to-use nano/microfibers obtainable by said method, as well as uses thereof, including wound healing, drug delivery, and (therapeutic) biomedical applications such as tissue regeneration and engineering. do.

Description

Method for preparing poly(alkyl cyanoacrylate)-based nano/microfiber and use thereof

The present invention relates to the field of biomaterials, and more particularly to the field of poly(alkyl cyanoacrylate) based nano- and microfibers. The present invention provides a novel process for the preparation of ready-to-use poly(alkyl cyanoacrylate)-based nano/microfibers, which process is produced by anionic polymerization of alkyl cyanoacrylate monomers or oligomers and has a specific polydispersity. electrospinning of poly(alkyl cyanoacrylate) homopolymers or copolymers characterized by a polydispersity index. Accordingly, the present invention also provides novel ready-to-use nano/microfibers obtainable by said method, as well as uses thereof, including (therapeutic) biomedical applications such as wound healing, drug delivery and tissue regeneration and tissue engineering. do.

Poly(alkyl cyanoacrylate) polymers (PACA polymers) can be produced by direct Knoevenagel condensation of each alkyl cyanoacetate with formaldehyde (Nicolas and Couvreur, 2009, Synthesis of Poly (alkyl cyanoacrylate )-based Colloidal Nanomedicines . Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol., Vol. 1(1): 111-127]). PACA polymers have been studied for use in controlled drug delivery systems, prepared by polymerization of alkyl cyanoacrylates (ACA), or by precipitation of preformed PACAs thereof from organic solvents.

Several techniques are known for producing PACA-based nanofibers, one of which is electrospinning (Teo et al., 2011, Technological Advances in Electrospinning of Nanofibers , Sci. Technol. Adv. Mater., Vol. 12 (1):013002, 19 pp.]). Briefly, electrospinning technology uses electrical force to draw a charged thread of a polymer solution on a collector to a fiber diameter in the sub-nanometer range. One of the simplest electrospinning machines consists of a high voltage power source, an infusion pump system, a syringe with a needle, and a collector. WO 2014/184761 relates to a method for producing polycyanoacrylate fibers using electrospinning, in a dipole aprotic solvent performing dual functions of a solvent for cyanoacrylate monomer and a catalyst for initiating polymerization. The formation of cyanoacrylate polymers of

PACA-based electro-spun nanofibers are considered biocompatible and form a scaffold of nanofiber meshes that can be used, for example, in a variety of biomedical applications. US 2014/0205971 A1 discloses the preparation of nanofibers by electrospinning of other synthetic polymers such as polycaprolactone (PCL) and poly(lactic-co-glycolic) acid (PLGA), and bone grafting, tissue in dental implants. Its use for growth and regeneration is described. Dental implants have become a part of everyday dentistry, and there is a growing interest in soft tissue healing around implants. However, to this day, dental implant surgery is still associated with complications such as material-related wound dehiscence.

There continues to be a need for new methods for making PACA-based nano/microfibers that can be used as scaffolds for novel ready-to-use products such as nanofiber meshes and their use in biomedical applications. Due to limitations in the art, particularly with regard to wound dressing-related complications in dental implant surgery, the present invention provides a novel method for preparing ready-to-use poly(alkyl cyanoacrylate)-based nano/microfibers, as well as the method Addresses the needs of the art by providing the nano/microfibers obtainable by

[Summary of the invention]

The present disclosure provides the following [1] to [15], but is not specifically limited thereto;

[1] A method for preparing a ready-to-use poly(alkyl cyanoacrylate)-based nano/microfiber comprising:

(a) poly(alkyl cyanoacrylate) homopolymers and/or poly(alkyl cyanoacrylates) obtained by anionic polymerization of at least one alkyl cyanoacrylate (ACA) monomer or at least one ACA oligomer providing a copolymer, wherein the alkyl substituents of the ACA monomer or ACA oligomer may be optionally substituted and the number-average molecular weight of the poly(alkyl cyanoacrylate) homopolymer or poly(alkyl cyanoacrylate) copolymer the ratio of the weight-average molecular weight (M _w ) to (M _n ) is from 1 to 1.7;

(b) dissolving the poly(alkyl cyanoacrylate) homopolymer or poly(alkyl cyanoacrylate) copolymer of step (a) in a solvent;

(c) electrospinning the solution obtained in step b) to obtain poly(alkyl cyanoacrylate)-based nano/microfibers on a collector; and

(d) removing the poly(alkyl cyanoacrylate)-based nano/microfibers from the collector to obtain ready-to-use poly(alkyl cyanoacrylate)-based nano/microfibers.

[2] the poly(alkyl cyanoacrylate) copolymer of step (a) comprises (i) at least two different ACA monomer species, (ii) at least two alkyl species comprising at least two different ACA monomer species The method of [1], comprising a cyanoacrylate oligomer, or (iii) at least one poly(alkyl cyanoacrylate) (PACA) polymer and at least one non-PACA polymer.

[3] at least one ACA monomer or at least two different ACA monomer species n-butyl-2-cyanoacrylate (BCA), iso-butyl-2-cyanoacrylate (IBCA), n-pentyl-2 -Cyanoacrylate (PCA), iso-pentyl-2-cyanoacrylate (IPCA), n-hexyl-2-cyanoacrylate (HCA), iso-hexyl-2-cyanoacrylate (IHCA) , Cyclo-hexyl-2-cyanoacrylate (CHCA), n-heptyl-2-cyanoacrylate (HepCA), iso-heptyl-2-cyanoacrylate (IHepCA), n-octyl-2-cya [1] or [ 2] method.

[4] at least one ACA oligomer is an oligomer of n-butyl-2-cyanoacrylate (BCA), an oligomer of iso-butyl-2-cyanoacrylate (IBCA), n-pentyl-2-cyanoacryl oligomer of rate (PCA), oligomer of iso-pentyl-2-cyanoacrylate (IPCA), oligomer of n-hexyl-2-cyanoacrylate (HCA), iso-hexyl-2-cyanoacrylate ( oligomer of IHCA), oligomer of cyclo-hexyl-2-cyanoacrylate (CHCA), oligomer of n-heptyl-2-cyanoacrylate (HepCA), iso-heptyl-2-cyanoacrylate (IHepCA) of oligomer, oligomer of n-octyl-2-cyanoacrylate (OCA), oligomer of iso-octyl-2-cyanoacrylate (IOCA), butyl-lactoyl-2-cyanoacrylate (BLCA) The method of [1] or [2], selected from any of oligomers and mixtures thereof.

[5] The solvent of step [b] is selected from any one of acetone, ethanol, acetic acid, formic acid, ethyl acetate, dimethyl sulfoxide, dimethyl formamide, butyl acetate, isobutyl acetate, and mixtures thereof [1] to [ 4] any of the methods.

[6] The method of any of [1] to [5], wherein at least one non-PACA polymer is added to the solution obtained in step (b) prior to electrospinning. In various embodiments, said at least one non-PACA polymer is at least one non-PACA homopolymer and/or at least one non-PACA copolymer.

[7] at least one non-PACA polymer is poly(lactic acid-co-glycolic acid) (PLGA), poly(ε-caprolactone) (PCL), poly(ethylene glycol) (PEG), poly(lactic acid) ( PLA), chitosan (CH), hyaluronic acid (HA), and dextran (Dex), the method of any one of [2] to [6].

[8] The method of any of [1] to [7], wherein the ready-to-use nano/microfibers obtained in step (d) are not included in the support.

[9] the ratio of the weight average molecular weight to the number average molecular weight is 1.1 to 1.5; or the method of any one of [1] to [8], wherein the ratio of the weight average molecular weight to the number average molecular weight is 1.2 to 1.4 or 1.4 to 1.7.

[10] Ready-to-use nano/microfibers obtainable by or obtained by the method of any one of [1] to [9].

[11] The ready-to-use nano/microfibers of [10], wherein the average fiber diameter is from 100 nm to 5 micrometers.

[12] Nano/microfibers are therapeutic agents, antibiotics, antiviral agents, chemotherapeutic agents, analgesic and analgesic combinations, anti-inflammatory agents, vitamins, sedatives, radiopharmaceuticals, metal particles, metal alloy particles, metal oxide particles, hydroxyapatite, sodium alginate , dyes, cells, proteins, peptides, nucleic acids, nucleic acid analogs, nucleotides or oligonucleotides, peptide nucleic acids, aptamers, antibodies, or fragments or portions thereof, antigens or epitopes, hormones, hormone antagonists, growth factors or recombinant growth factors, and The ready-to-use nano/microfibers of [10], wherein they are immobilized or encapsulated with at least one of fragments and variants thereof, cell adhesion mediators, cytokines, enzymes or mixtures thereof.

[13] The ready-to-use nano/microfiber of [10], wherein the nano/microfiber additionally comprises a cell growth medium.

[14] Use of the ready-to-use nano/microfibers according to [10] in dental applications, for wound dressing/healing, tissue regeneration/engineering, organ protection, implant coating or controlled drug delivery.

[15] A nanofiber mesh comprising the ready-to-use nano/microfibers of any of [10] to [13], wherein the surface of the nanofiber mesh is chemically and/or physically modified for biomedical applications, Nanofiber mesh, preferably wherein the surface is modified by encapsulation and/or immobilization of therapeutic agents and/or biological materials.

Additional aspects and embodiments of the invention are set forth in the detailed description that follows.

1 shows a representative electrospinning apparatus. A) High voltage power supply, B) Injection system pump, C) Ground-connected collector.
2 shows a representative infusion system pump in which the syringe is associated with the needle and the collector is grounded.
3 shows an electro-spun mesh of PACA substrate collected on a copper mesh used as a collector.
4 shows an electro-spun mesh of PACA substrate collected on different collectors. A: copper ring, B: aluminum foil, C: paper.
FIG. 5 shows fluorescence photographs at different magnifications of human gingival fibroblasts (HGFib) grown on PBCA meshes for 14 days of incubation at 37° C. and 5% (V/V) CO _{2 .} A: After 3 days of incubation, B: After 7 days of incubation, C: After 14 days of incubation. Each magnification of AC: left bar 500 nm, middle bar 200 nm, right bar 100 nm.
6 shows SEM (Scanning Electron Microscopy) pictures of meshes based on PACA. A and B: PACA with different molecular weight, rod = 5 μm in two pictures of A, B: rod 5 μm in top two pictures, rod 2 μm in bottom two pictures, bar 10 μm in middle left picture , 5 μm bar in the middle right photo; C: PACA/PLGA; D: PACA/5FU; E: PACA/PACA-co-CS.
7 shows A: powder of PACA; B: powder of PACA-co-CS; C: GPC (gel permeation chromatography) results of electro-spun meshes of PACA/PACA-co-CS are shown.
8 depicts the procedure for seeding cells onto an electro-spun PACA mesh. A: UV irradiation; B: addition of human gingival fibroblasts; C: Addition of cell culture medium.
9 shows the release of 5FU from PACA/5FU mesh incubated in PBS, pH 7.4 at 37°C. Three different samples were tested, m1, m2, m3, with different starting weights of 5FU encapsulated in each mesh: m1 = 7.30 mg, m2 = 6.67 mg, m3 = 6.12 mg. Samples were taken at different locations on the same PACA/5FU mesh prepared. 9 shows consistent drug release patterns up to 51-55% within 30 days in PBS medium. At day 96, drug release was 80-88% of total drug content in all samples.
10 is an SEM photograph of PBCA mesh before (A) and after 23 days (B) incubation with HGFib. The mesh was washed with water.
11 is a SEM photograph of meshes based on PACA (A2 and B2) at year 0 after manufacture (A1 and B1) and after one year (1 year) of storage under normal conditions (A2 and B2). A1 and A2: flat fibers; B1 and B2: circular fibers. Normal conditions: ambient temperature and pressure (25 ° C, 1 atmosphere).
12 is a SEM photograph of PLGA- and PACA/PLGA-based meshes at year 0 after fabrication (A1 and B1) and after one year (1 year) of storage under normal conditions (A2 and B2). Normal conditions: ambient temperature and pressure (25 ° C, 1 atmosphere).
13 shows confocal laser scanning microscopy of PACA mesh stained with D&C Violet #2.
14 shows PBCA meshes with fiber diameters ranging from 0.1 to 5 μm: (A) 4.049±2.087 (B) 1.866±0.799 μm (C) 1.232±0.544 μm (D) 0.685±0.569 μm (E) 1.6± 0.5 μm (F) 0.8±0.1 μm (G) 0.5±0.1 μm (H) 0.148±0.111 μm.

As the present invention, it is demonstrated that a mesh or mesh network of PACA nano/microfibers can be produced by electrospinning of a PACA solution. Different types of meshes were made as shown in the examples and accompanying drawings. The present invention provides as a general concept a process for the production of poly(alkyl cyanoacrylate) (PACA)-based nano/microfibers and/or PACA-based nano/microfiber meshes, comprising:

(a) providing a poly(alkyl cyanoacrylate) homopolymer and/or a poly(alkyl cyanoacrylate) copolymer, wherein the poly(alkyl cyanoacrylate) homopolymer and/or poly(alkyl cyano the ratio of the weight-average molecular weight (M _w ) to the number-average molecular weight (M _n ) of the noacrylate) copolymer is 1 to 1.7;

(b) dissolving the poly(alkyl cyanoacrylate) homopolymer or poly(alkyl cyanoacrylate) copolymer of step (a) in a solvent; and

(c) electrospinning the solution obtained in step (b) to obtain poly(alkyl cyanoacrylate)-based nano/microfibers, preferably electrospinning the solution obtained in step (b) to obtain a collector Obtaining a poly(alkyl cyanoacrylate)-based nano/microfiber on the phase.

In various embodiments, the alkyl substituents of the poly(alkyl cyanoacrylate) homopolymers and/or poly(alkyl cyanoacrylate) copolymers may be optionally substituted according to substitutions described elsewhere herein. Further, in various embodiments, the poly(alkyl cyanoacrylate) homopolymer and/or poly(alkyl cyanoacrylate) copolymer of step (a) comprises at least one alkyl cyanoacrylate (ACA) monomer and/or or obtainable by or obtained by ionic polymerization of one or more ACA oligomers, wherein the alkyl substituents of the ACA monomers and/or ACA oligomers may be optionally substituted. In various embodiments, the ionic polymerization is an anionic polymerization, or a cationic polymerization. Preferably, the ionic polymerization is anionic polymerization. The method includes an additional step of taking poly(alkyl cyanoacrylate) based nano/microfibers from the collector to obtain poly(alkyl cyanoacrylate) based nano/microfibers and/or PACA based nano/microfiber meshes ( d) may be included.

In various aspects, the method of making poly(alkyl cyanoacrylate)-based nano/microfibers comprises a poly(alkyl cyanoacrylate)-based nano/microfiber and/or PACA-based nano/microfiber mesh comprising is a manufacturing method of:

(a) poly(alkyl cyanoacrylate) homopolymers and/or poly(alkyl cyanoacrylics) obtained by ionic polymerization of at least one alkyl cyanoacrylate (ACA) monomer and/or at least one ACA oligomer rate) copolymer, wherein the alkyl substituents of the ACA monomers and/or ACA oligomers may be optionally substituted, the poly(alkyl cyanoacrylate) homopolymer or the poly(alkyl cyanoacrylate) copolymer the ratio of the weight-average molecular weight (M _w ) to the number-average molecular weight (M _n ) is 1 to 1.7;

(b) dissolving the poly(alkyl cyanoacrylate) homopolymer or poly(alkyl cyanoacrylate) copolymer of step (a) in a solvent;

(c) electrospinning the solution obtained in step b) to obtain a poly(alkyl cyanoacrylate)-based nano/microfiber mesh on a collector; and

(d) taking the poly(alkyl cyanoacrylate) based nano/microfiber mesh from the collector to obtain poly(alkyl cyanoacrylate) based nano/microfiber and/or PACA based nano/microfiber mesh.

In various embodiments, the ionic polymerization of step (a) is an anionic polymerization, or a cationic polymerization. Preferably, the ionic polymerization is anionic polymerization.

In various embodiments, PACA-based nano/microfiber meshes of the present invention may be described as PACA-based nano/microfiber networks or PACA-based nano/microfiber mesh networks. The terms may be used interchangeably herein. The term "mesh" (or "network" or "network mesh"), also known as a scaffold or mat, refers to those that can be invaded and colonized by cells, or acting as a protective barrier, in healing. to impart or provide an excellent environment, to provide a high surface area-to-volume ratio, to enable the exudation of fluid from a wound, to enable gas permeation, to inhibit the invasion of exogenous microorganisms into the wound fully-interconnected, which may have a variety of functions or properties, such as helping to regulate fluid drainage, and acting as a delivery platform for any kind of agent, including therapeutics, other chemicals, compounds, or biological materials. It may be considered to relate to a three-dimensional frame that is porous or even highly porous, including pores.

As used herein, the term “PACA” is used as an abbreviation for the term “poly(alkyl cyanoacrylate)”. Likewise, as used herein, the term “ACA” is used as an abbreviation for the term “alkyl cyanoacrylate”.

Also, the term “PACA-based nano/microfiber” encompasses “PACA-based nanofiber and/or PACA-based microfiber”. Likewise, the term “PACA-based nano/microfiber mesh(s)” encompasses “PACA-based nanofiber mesh(s)” and/or “PACA-based microfiber mesh(s)”.

As used herein, the term “PACA-based nanofiber(s)” means that the PACA-based fibers have an average diameter in the nanometer size range. In various aspects, "PACA-based nanofiber(s)" means that the nanofiber(s) have (average) diameters ranging from about 1 nm to about 999 nm, including (average) diameters ranging from 0.01 nm to 1 nm. means that In various embodiments, the “PACA-based nanofiber(s)” have a (average) diameter in the range of from about 100 nm to about 900 nm. In various other embodiments, the “PACA-based nanofiber(s)” have a (average) diameter in the range of about 200 nm to about 800 nm. In other embodiments, the “PACA-based nanofiber(s)” have a (average) diameter in the range from about 300 nm to about 700 nm. In another embodiment, the “PACA-based nanofiber(s)” have a (average) diameter in the range of from about 400 nm to about 600 nm. In various embodiments, the “PACA-based nanofiber(s)” has a (average) diameter of about 650 ± 200 nm or about 700 ± 230 nm. In various other embodiments, the “PACA-based nanofiber(s)” have a (average) diameter of about 675 ± 230 nm. In another embodiment, the “PACA-based nanofiber(s)” has a (average) diameter of about 700 ± 200 nm or about 700 ± 230 nm.

Likewise, as used herein, the term “PACA-based microfiber(s)” means that the PACA-based fibers have an average diameter in the micrometer size range. In various aspects, "PACA-based microfiber(s)" means that the microfiber(s) have (average) diameters ranging from about 1 μm to about 1,000 μm, including (average) diameters ranging between 0.01 μm to 1 μm. means that In various embodiments, the “PACA-based microfiber(s)” have a (average) diameter ranging from about 100 μm to about 900 μm. In various other embodiments, the “PACA-based microfiber(s)” have a (average) diameter ranging from about 200 μm to about 800 μm. In another embodiment, the “PACA-based microfiber(s)” have a (average) diameter in the range of from about 300 μm to about 700 μm. In another embodiment, the “PACA-based microfiber(s)” have a (average) diameter in the range of about 400 μm to about 600 μm. In various embodiments, the “PACA-based microfiber(s)” have a (average) diameter of about 500 ± 200 μm or about 500 ± 250 nm. In various other embodiments, the “PACA-based microfiber(s)” have a (average) diameter of about 10 ± 5 μm. In another embodiment, the “PACA-based microfiber(s)” have a (average) diameter of about 2 ± 1 μm.

In various embodiments, “PACA-based nano/microfiber(s)” are in the range of about 600 nm to about 1,400 nm, preferably in the range of about 700 nm to about 1,300 nm, more preferably in the range of about 800 nm to about 1,200 nm. , even more preferably having a (average) diameter in the range from about 900 nm to about 1,100 nm. In various embodiments, the “PACA-based nano/microfiber(s)” have a (average) diameter in the range of about 1,000±400 nm or in the range of about 1,000±250 nm. In various other embodiments, “PACA-based nano/microfiber(s)” are in the range of about 1,000 ± 300 nm, preferably in the range of about 1,000 ± 250 nm, more preferably in the range of about 1,000 ± 200 nm or even about 1,000 It has a (average) diameter in the range ± 100 nm.

In various embodiments, the PACA-based nano/microfibers have an average diameter in the range of about 100 nm to 5 μm. In various preferred embodiments, the PACA-based nano/microfibers may have an average diameter ranging from about 100 nm to 200 nm or from 100 nm to 250 nm. In various other preferred embodiments, the PACA-based nano/microfibers may have an average diameter ranging from about 1.4 μm to about 5 μm. In another preferred embodiment, the PACA-based nano/microfibers may have an average diameter ranging from about 1.4 μm to about 3.2 μm. In another preferred embodiment, the PACA-based nano/microfibers may have an average diameter in the range of about 1.4 μm to 2 μm. In another preferred embodiment, the PACA-based nano/microfibers may have an average diameter ranging from about 3.2 μm to 5 μm or from about 2 μm to 5 μm. In particularly preferred embodiments, the PACA-based nano/microfibers have an average diameter of about 700 nm or about 1 μm or about 1.3 μm. Accordingly, in another particularly preferred embodiment, the PACA-based nano/microfibers have an average diameter in the range of about 700 nm to about 1.3 μm, or in the range of about 700 nm to about 1 μm, or in the range of about 1 μm to about 1.3 μm. .

In various aspects, a method for producing a PACA-based nano/microfiber or a PACA-based nano/microfiber mesh may each include step (a) as follows:

(a) poly(alkyl cyanoacrylate) homopolymers and/or poly(alkyl cyanoacrylates) by ionic polymerization of one or more alkyl cyanoacrylate (ACA) monomers and/or one or more ACA oligomers producing (or obtaining) a copolymer, wherein the alkyl substituents of the ACA monomers and/or ACA oligomers may be optionally substituted, poly(alkyl cyanoacrylate) homopolymers and/or poly(alkyl cyanoacrylics) rate) the ratio of the weight-average molecular weight (M _w ) to the number-average molecular weight (M _n ) of the copolymer is from 1 to 1.7. The following step (b) is thus represented as follows: (b) the poly(alkyl cyanoacrylate) homopolymer and/or poly(alkyl cyanoacrylate) produced (or obtained) in step (a) ) dissolving the copolymer in a solvent. In various embodiments, the ionic polymerization is an anionic polymerization, or a cationic polymerization. Preferably, the ionic polymerization is anionic polymerization.

In step (b) of the process according to the invention, any suitable solvent can be used as long as the poly(alkyl cyanoacrylate) homopolymer or poly(alkyl cyanoacrylate) copolymer is sufficiently dissolved therein. A solution is obtained by dissolving the PACA homopolymer or PACA copolymer therein, preferably by mixing using magnetic stirring until the homopolymer or copolymer is completely dissolved to obtain a clear solution. When more than one polymer is used for electrospinning (such as when a non-PACA polymer is added to the solution obtained in step (b) of the method described herein prior to electrospinning), each polymer solution is prepared separately After that, their mixing can follow. Alternatively, a solution comprising only one PACA homopolymer or PACA copolymer may be prepared followed by addition of additional components.

In various embodiments of the invention, the poly(alkyl cyanoacrylate) homopolymer and/or poly(alkyl cyanoacrylate) copolymer is a (direct) brain between the corresponding alkyl cyanoacetate(s) and formaldehyde. obtainable by Venagel condensation, and/or ionic polymerization of one or more ACA monomers and/or one or more ACA oligomers, and/or ionic polymerization of one or more ACA monomers with one or more non-PACA polymers. may be, or may be obtained, or may be produced. The non-PACA polymer is capable of catalyzing or initiating the ionic polymerization of an ACA monomer due to the ionic group of the non-PACA polymer, ie, the ionic group acts as an initiator of the anionic polymerization of the ACA monomer. In various embodiments, the aforementioned ionic polymerization is anionic polymerization, or cationic polymerization. Preferably, the ionic polymerization is anionic polymerization. In accordance with the foregoing, the process described herein comprises step (a) (i) a (direct) Neubenagel condensation between the corresponding alkyl cyanoacetate(s) and formaldehyde, and/or (ii) one or more ionic (anionic or cationic) polymerization of ACA monomers and/or one or more ACA oligomers, and/or (iii) ionic (anionic or cationic) polymerization of one or more ACA monomers with one or more non-PACA polymers. sex) polymerization.

Also disclosed herein is a method of making (ready-to-use) poly(alkyl cyanoacrylate)-based nano/microfibers and/or PACA-based nano/microfiber meshes comprising:

(a) providing a poly(alkyl cyanoacrylate) homopolymer and/or a poly(alkyl cyanoacrylate) copolymer, wherein the poly(alkyl cyanoacrylate) homopolymer and/or poly(alkyl cyano the ratio of the weight-average molecular weight (M _w ) to the number-average molecular weight (M _n ) of the noacrylate) copolymer is 1 to 1.7;

(b) dissolving the poly(alkyl cyanoacrylate) homopolymer and/or poly(alkyl cyanoacrylate) copolymer of step (a) in a solvent; and

(c) electrospinning the solution obtained in step (b) to obtain poly(alkyl cyanoacrylate) based nano/microfiber and/or PACA based nano/microfiber mesh, preferably step (b) ) to obtain a poly(alkyl cyanoacrylate)-based nano/microfiber and/or PACA-based nano/microfiber mesh on a collector by electrospinning the solution obtained.

The above-mentioned method takes out the poly(alkyl cyanoacrylate) based nano/microfiber and/or PACA based nano/microfiber mesh from the collector and provides ready-to-use poly(alkyl cyanoacrylate) based nano/microfiber mesh. and/or an additional step (d) of obtaining a PACA-based nano/microfiber mesh. In various embodiments, the poly(alkyl cyanoacrylate) copolymer comprises or is formed by (i) at least one PACA polymer, and (ii) at least one non-PACA polymer. The non-PACA polymer may be any non-PACA polymer as described elsewhere herein. Likewise, the PACA polymer may be any PACA polymer as described elsewhere herein. In various embodiments, the poly(alkyl cyanoacrylate) copolymer comprising or formed by (i) at least one PACA polymer, and (ii) at least one non-PACA polymer comprises (i) 1 Obtainable by, obtainable by, or produced by the ionic polymerization of at least one alkyl cyanoacrylate (ACA) monomer (or ACA oligomer) with (ii) at least one non-PACA polymer. The ionic polymerization may be an anionic polymerization, or it may be a cationic polymerization. Preferably, the ionic polymerization is anionic polymerization. The alkyl substituents of the ACA monomers and/or ACA oligomers may be optionally substituted. In various embodiments, ionic (anionic or cationic) polymerization may be initiated by an initiator, which may be an initiator as described elsewhere herein. In various preferred embodiments, the ionic (anionic or cationic) polymerization is initiated by at least one non-PACA polymer. Surprisingly, the inventors have found that ACA monomers can be polymerized using the ionic groups of non-PACA polymers as initiators for polymerization. This makes it possible to prepare copolymers, which may also be referred to as graft copolymers. The present disclosure particularly relates to (ready-to-use) PACA-based nano/microfibers (and/or PACA-based nanofibers) obtainable by, or obtained by, a process for preparing the corresponding grafted copolymer as described herein /microfiber mesh).

Also disclosed herein is a process for making (ready-to-use) poly(alkyl cyanoacrylate)-based nano/microfibers and/or PACA-based nano/microfiber meshes comprising the steps of:

(a) providing a poly(alkyl cyanoacrylate) copolymer comprising or formed by a PACA polymer and a non-PACA polymer;

(b) dissolving the poly(alkyl cyanoacrylate) copolymer of step (a) in a solvent; and

(c) electrospinning the solution obtained in step (b) to obtain poly(alkyl cyanoacrylate) based nano/microfiber and/or PACA based nano/microfiber mesh, preferably step (b) ) to obtain a poly(alkyl cyanoacrylate)-based nano/microfiber and/or PACA-based nano/microfiber mesh on a collector by electrospinning the solution obtained.

In a preferred embodiment, the ratio of the weight-average molecular weight (M _{w ) to the number-average molecular weight (M n} ) of the poly(alkyl cyanoacrylate) copolymer is from 1 to 1.7. The method takes poly(alkyl cyanoacrylate)-based nano/microfibers and/or PACA-based nano/microfiber meshes from the collector, ready-to-use poly(alkyl cyanoacrylate)-based nano/microfibers and/or or an additional step (d) of obtaining a PACA-based nano/microfiber mesh. In various embodiments, the poly(alkyl cyanoacrylate) copolymer comprises or is formed by (i) at least one PACA polymer, and (ii) at least one non-PACA polymer. In a preferred embodiment, the PACA copolymer comprises (i) at least one PACA homopolymer, and (ii) at least one non-PACA polymer. The non-PACA polymer can be any non-PACA polymer as described anywhere herein, including any non-PACA homopolymer and any non-PACA copolymer described anywhere herein. . Likewise, the PACA (homo)polymer may be any PACA (homo)polymer as described elsewhere herein. In various embodiments, the PACA copolymer comprising or formed by at least one PACA (homo)polymer and at least one non-PACA polymer comprises at least one alkyl cyanoacrylate (ACA) monomer (or ACA oligomer). ) and at least one non-PACA polymer obtainable by, obtainable by, or produced thereby. The ionic polymerization may be an anionic polymerization, or it may be a cationic polymerization. Preferably, the ionic polymerization is anionic polymerization. The alkyl substituents of the ACA monomers and/or ACA oligomers may be optionally substituted. In various embodiments, the ionic polymerization may be initiated by an initiator, which may be an initiator as described elsewhere herein. In various preferred embodiments, the ionic (anionic or cationic) polymerization is initiated by at least one non-PACA polymer. Surprisingly, the inventors have found that ACA monomers can be polymerized using the ionic groups of non-PACA polymers as initiators for polymerization. This makes it possible to prepare copolymers, which may also be referred to as graft copolymers. The present disclosure particularly relates to (ready-to-use) PACA-based nano/microfibers (and/or PACA-based nanofibers) obtainable by, or obtained by, a process for preparing the corresponding grafted copolymer as described herein /microfiber mesh).

In various embodiments, the PACA-based nano/microfiber or PACA-based nano/microfiber mesh produced is a ready-to-use PACA-based nano/microfiber or a ready-to-use PACA-based nano/microfiber mesh, respectively. In various embodiments, the prepared PACA-based nano/microfibers or PACA-based nano/microfiber meshes are ready-to-use PACA-based nano/microfibers or ready-to-use PACA-based nano/microfiber meshes, respectively, comprising at least one This is because the PACA homopolymer and/or the one or more PACA copolymers are dissolved in a solvent that is biocompatible (see step (b) of the method above). In various preferred embodiments, said biocompatible solvent is biocompatible with skin, bone and/or tissue. The term “skin”, “bone” or “tissue” as used herein encompasses animal and human skin, bone or tissue. In various embodiments, the term "biocompatible solvent" refers to solvents known to be used in the manufacture of drug substances, excipients, and drug products because the solvent is a non-toxic solvent and/or the solvent does not have any deleterious environmental effects. it means. Accordingly, in various embodiments, a "biocompatible solvent" is a non-toxic solvent and/or does not have any deleterious environmental effects. Preferably, the solvent used to dissolve the at least one PACA homopolymer and/or the at least one PACA copolymer is a solvent that must not be used in the manufacture of drug substances, excipients and drug products (“Class 1 solvents”). "), and compared to solvents that must be restricted in pharmaceutical products due to their intrinsic toxicity ("Class 2 solvents"), and may be considered to have a lower risk to human health. Accordingly, in the present invention, the solvent used to dissolve the one or more PACA (homo)polymers and/or the one or more PACA copolymers is a "class 3 solvent", i.e. compared to a solvent of class 1 or 2 It is a solvent that is less toxic when used and can be considered as having a lower risk to human health. Solvents of classes 1, 2 and 3 are known to the person skilled in the art, for example, from industry guidelines provided by the US FDA (Food and Drug Administration). In particular, see Human Use guidance for industry " Q3C Impurities: Residual Solvents" (Revision 3 of June 2017), which makes recommendations on what amounts of residual solvents are considered safe in pharmaceuticals. "Class 3" solvents include acetic acid, acetone, anisole, 1-butanol, 2-butanol, butyl acetate, tert -butylmethyl ether, dimethyl sulfoxide (DMSO), ethanol, ethyl acetate, ethyl ether, ethyl formate, Formic acid, heptane, isobutyl acetate, isopropyl acetate, methyl acetate, 3-methyl-1-butanol, methylethyl ketone, 2-methyl-1-propanol, pentane, 1-pentanol, 1-propanol, propyl acetate, trimethyl amines, dimethyl formamide, and any combination or mixture thereof. Any of these solvents may be used to dissolve one or more PACA (homo)polymers and/or one or more PACA copolymers according to the process of the present invention. In various embodiments, the solvent is selected from any of acetone, ethanol, acetic acid, formic acid, ethyl acetate, dimethyl sulfoxide (DMSO), dimethyl formamide, butyl acetate, isobutyl acetate, and any combination or mixture thereof. In a particularly preferred embodiment, the solvent is acetone. Such an embodiment has been shown to produce an average fiber diameter of about 2 μm using about 1% polymer solution. In various other preferred embodiments, the solvent is an acetone:ethanol mixture. Preferably, the solvent is an acetone:ethanol mixture of about 4:1 (v/v). Such an embodiment has been shown to produce an average fiber diameter of about 700±100 nm using about 1% polymer solution.

U.S. The following solvents for which no toxicological data have been found according to the FDA may be used in the context of this disclosure: 1,1-diethoxypropane, 1,1-dimethoxymethane, 2,2-dimethoxypropane, isooctane, Any of isopropyl ether, methyl isopropyl ketone, methyl tetrahydrofuran, petroleum ether, trichloroacetic acid and trifluoroacetic acid. In various preferred embodiments, the solvent is trichloroacetic acid or trifluoroacetic acid or mixtures thereof. In various preferred embodiments, the solvent is a mixture of acetone:trichloroacetic acid or acetone:trifluoroacetic acid. Such mixtures may be used to prepare blend solutions of PACA and non-PACA polymers as described herein, particularly blend solutions of PACA and chitosan or PACA and PCL. Preferably, the mixture of acetone:trichloroacetic acid or acetone:trifluoroacetic acid is a mixture of about 4:1 (v/v).

In accordance with the above, the PACA-based nano/microfibers or PACA-based nano/microfiber meshes provided by the present invention may be considered to be suitably biocompatible, in particular biocompatible with skin or tissue. The term "skin" or "tissue" as used herein encompasses animal and human skin or tissue. Said PACA-based nano/microfiber or PACA-based nano/microfiber mesh can be considered biocompatible, as it is non-toxic and/or does not have any deleterious effect on the human or animal body, thus (therapeutic ) as it may be used in biomedical applications and/or drug products.

Advantageously, the process according to the invention provides a PACA-based nano/microfiber or a PACA-based nano/microfiber mesh that does not require additional manufacturing steps to obtain a ready-to-use product, for example a ready-to-use product. . Thus, ready-to-use nano/microfibers or ready-to-use nano/microfiber meshes according to the invention preferably do not comprise or require no support. Advantageously, biodegradable nano/microfibers or nano/microfiber meshes can be used for a variety of different biomedical applications, including therapeutic biomedical applications, and can degrade over time without the need for a support. In particular, the biodegradable nano/microfibers or nano/microfiber meshes of the invention can advantageously be used in dental applications, including, for example, dental surgery, in particular oral and maxillofacial surgery. Furthermore, there are no concerns regarding the compatibility of the support material with the nano/microfiber or nano/microfiber mesh material(s). Moreover, when used in medical applications, there are no concerns regarding any acceptability issues of the support material with respect to the patient to be treated. Accordingly, in various embodiments, since the nano/microfibers are not included in the support, the prepared PACA-based nano/microfibers or PACA-based nano/microfiber meshes are respectively ready-to-use PACA-based nano/microfibers or ready-to-use -Available PACA-based nano/microfiber mesh. Specifically, in various embodiments, after removal of the PACA-based nano/microfiber or PACA-based nano/microfiber mesh from the collector to obtain a ready-to-use PACA-based nano/microfiber or PACA-based nano/microfiber mesh, respectively , The method for preparing a ready-to-use PACA-based nano/microfiber or ready-to-use PACA-based nano/microfiber mesh comprises providing a PACA-based nano/microfiber or PACA-based nano/microfiber mesh on a support. No subsequent steps are involved.

In the present invention, the (alkyl) a cyanoacrylate component, of at least one, such as is represented by the formula _{H 2 C = C (CN)} -COOR , where R is is not limited to include any of the C _1-15 alkyl group ( alkyl) cyanoacrylate monomers. The alkyl chain may be a linear chain or a branched chain C _1-15 alkyl chain. Specifically, the at least one cyanoacrylate monomer of the present disclosure _{may be represented by the formula H 2} C=C(CN)-COOR, wherein R is a C ₁ alkyl chain, a C ₂ alkyl chain, C ₃ alkyl chain, C ₄ alkyl chain, C ₅ alkyl chain, C ₆ alkyl chain, C ₇ alkyl chain, C ₈ alkyl chain, C ₉ alkyl chain, C ₁₀ alkyl chain, C ₁₁ alkyl chain, C ₁₂ alkyl chain, C ₁₃ an alkyl chain, a C ₁₄ alkyl chain, a C ₁₅ alkyl chain, and mixtures thereof. Preferably, the alkyl chain is a straight or branched C _1-10 alkyl chain, more preferably the alkyl chain is a straight or branched C _1-8 alkyl chain.

Accordingly, in various aspects of the invention, the alkyl substituents of the ACA monomers have a chain length of from 1 to 15 carbon atoms, preferably from 1 to 10 carbon atoms, more preferably from 1 to 8 carbon atoms. chain length. In various preferred embodiments, the alkyl chain is a linear C _1-10 alkyl chain or a branched C _1-10 alkyl chain. More preferably, the alkyl chain is a linear C _1-8 alkyl chain or a branched C _1-8 alkyl chain. Even more preferably, the alkyl chain is a linear C _1-4 alkyl chain or a branched C _1-4 alkyl chain. Even more preferably, the alkyl chain is a linear C _1-3 alkyl chain or a branched C _1-3 alkyl chain. In a particularly preferred embodiment, the alkyl chain is a linear C _1-2 alkyl chain or a branched C _1-2 alkyl chain. Preferably, the linear alkyl chain is a linear straight chain alkyl chain.

In accordance with the above, in various aspects of the present invention, the alkyl substituents of each monomer unit constituting the ACA oligomers as described herein have a chain length of 1 to 15 carbon atoms, preferably a chain of 1 to 10 carbon atoms. have a length Preferably, the alkyl chain of each monomer unit constituting the at least one or more ACA oligomers is a linear C _1-10 alkyl chain or a branched C _1-10 alkyl chain. More preferably, the alkyl chain of each monomer unit constituting the at least one or more ACA oligomers is a linear C _1-8 alkyl chain or a branched C _1-8 alkyl chain. Even more preferably, the alkyl chain of each monomer unit constituting the at least one or more ACA oligomers is a linear C _1-5 alkyl chain or a branched C _1-5 alkyl chain. Even more preferably, the alkyl chain of each monomer unit constituting the at least one or more ACA oligomers is a linear C _1-3 alkyl chain or a branched C _1-3 alkyl chain. In a particularly preferred embodiment, the alkyl chain of each monomer unit constituting the at least one or more ACA oligomers is a linear C _1-2 alkyl chain or a branched C _1-2 alkyl chain. Preferably, said linear alkyl chain of each monomer unit constituting at least one or more ACA oligomers is a linear linear alkyl chain.

Branched alkanes can be derived from (linear) straight chain alkane systems by removing one of the hydrogen atoms from a methylene group (-CH2-) and replacing it with an alkyl group (thus the alkyl group becomes a substituent). Accordingly, in various embodiments of the present disclosure, at least one methylene group (-CH2-) of a branched alkyl chain disclosed herein is substituted by an alkyl group substituent. Accordingly, in various embodiments, the branched alkyl chains described herein are characterized by an alkyl group substituent attached to the alkyl chain. The alkyl group substituent bonded to the alkyl chain of at least one cyanoacrylate monomer may be any _{of a C 1-15 alkyl group.} Preferably, the alkyl group substituent attached to the alkyl chain of at least one cyanoacrylate monomer _{is any of a C 1-10} alkyl group. More preferably, the alkyl group bonded to the alkyl chain of at least one cyanoacrylate monomer is any of _{C 1-8 alkyl groups.} Even more preferably, the alkyl group substituent attached to the alkyl chain of at least one cyanoacrylate monomer _{is any of a C 1-4} alkyl group.

Likewise, the alkyl group substituent bonded to the alkyl chain of each monomer unit constituting at least one or more ACA oligomers may be any _{of C 1-15 alkyl groups.} Preferably, the alkyl group substituent bonded to the alkyl chain of each monomer unit constituting the at least one or more ACA oligomers _{is any of C 1-10} alkyl groups. More preferably, the alkyl group substituent bonded to the alkyl chain of each monomer unit constituting the at least one or more ACA oligomers _{is any of C 1-8} alkyl groups, even more preferably any of C _1-4 alkyl groups. . The alkyl group substituent may include a heteroatom and a functional group, such as a hydroxy group, a substituent including an ether linkage, an amino group, a carboxy group, and mixtures thereof. A more preferred substituent comprising a heteroatom is exemplified by the monomer butyl-lactoyl-2-cyanoacrylate (BLCA). In one preferred embodiment, the substituents contain only carbon and hydrogen atoms.

The alkyl group bonded to the alkyl chain of at least one cyanoacrylate monomer may be a straight chain or branched chain alkyl group. Preferably, the alkyl group bonded to the alkyl chain of at least one cyanoacrylate monomer is a straight chain alkyl group.

Likewise, in various embodiments of the present disclosure, the alkyl chain of at least one cyanoacrylate monomer is a linear alkyl chain.

Similar to the previous description, in this disclosure, the poly(alkyl cyanoacrylate) prepared by polymerization may be considered a (linear) straight-chain type or branched-chain type poly(alkyl cyanoacrylate) polymer.

In various embodiments, the (alkyl) cyanoacrylate monomer or monomer unit is methyl cyanoacrylate, 2-methoxy ethyl cyanoacrylate, ethyl cyanoacrylate, propyl cyanoacrylate, butyl cyanoacrylic rate (such as n -butyl 2-cyanoacrylate), hexyl cyanoacrylate, isohexyl (2-)cyanoacrylate, octyl cyanoacrylate, (2-)octyl cyanoacrylate, isobutyl ( 2-) at least one of cyanoacrylates and combinations thereof. In a preferred embodiment, the (alkyl) cyanoacrylate monomer or monomer unit is methyl cyanoacrylate, ethyl cyanoacrylate, butyl cyanoacrylate (such as n -butyl 2-cyanoacrylate), octyl cyano at least one of acrylates and combinations thereof. In various other preferred embodiments, the (alkyl) cyanoacrylate monomer or monomer unit is selected from at least one of methyl cyanoacrylate, ethyl cyanoacrylate, octyl cyanoacrylate, and combinations thereof. In another preferred embodiment, the (alkyl) cyanoacrylate monomer is selected from at least one of methyl cyanoacrylate, ethyl cyanoacrylate, butyl cyanoacrylate and combinations thereof. In various other embodiments, the cyanoacrylate monomer is selected from ethyl cyanoacrylate, butyl cyanoacrylate, and combinations thereof. A particularly preferred cyanoacrylate monomer or monomer unit includes ethyl cyanoacrylate. Suitably, the cyanoacrylate monomer or monomer unit may be ethyl cyanoacrylate. Another particularly preferred cyanoacrylate monomer or monomer unit is butyl cyanoacrylate (eg n -butyl 2-cyanoacrylate). The aforementioned ACA monomers or monomer units may form the basis of the ACA oligomers as described herein. Accordingly, in various embodiments of the present invention, said at least one ACA oligomer is selected from oligomers of any of the aforementioned ACA monomers or monomer units.

In various embodiments of the above method for making (ready-to-use) PACA-based nano/microfibers or PACA-based nano/microfiber meshes, one or more ACA monomers or at least two different ACA monomer species are n-butyl -2-cyanoacrylate (BCA), iso-butyl-2-cyanoacrylate (IBCA), n-pentyl-2-cyanoacrylate (PCA), iso-pentyl-2-cyanoacrylate ( IPCA), n-hexyl-2-cyanoacrylate (HCA), iso-hexyl-2-cyanoacrylate (IHCA), cyclo-hexyl-2-cyanoacrylate (CHCA), n-heptyl-2 -Cyanoacrylate (HepCA), iso-heptyl-2-cyanoacrylate (IHepCA), n-octyl-2-cyanoacrylate (OCA), iso-octyl-2-cyanoacrylate (IOCA) , butyl-lactoyl-2-cyanoacrylate (BLCA), and any combination or mixture thereof. One or more ACA monomers mentioned above may form the basis of an ACA oligomer as described herein. Thus, in various embodiments of the present invention, said one or more ACA oligomers are selected from oligomers of any of the aforementioned one or more ACA monomers. Thus, in various embodiments of the method of making (ready-to-use) PACA-based nano/microfibers or PACA-based nano/microfiber meshes, the at least one ACA oligomer comprises n-butyl-2-cyanoacrylate ( oligomer of BCA), oligomer of iso-butyl-2-cyanoacrylate (IBCA), oligomer of n-pentyl-2-cyanoacrylate (PCA), iso-pentyl-2-cyanoacrylate (IPCA) oligomer of, oligomer of n-hexyl-2-cyanoacrylate (HCA), oligomer of iso-hexyl-2-cyanoacrylate (IHCA), oligomer of cyclo-hexyl-2-cyanoacrylate (CHCA) , oligomer of n-heptyl-2-cyanoacrylate (HepCA), oligomer of iso-heptyl-2-cyanoacrylate (IHepCA), oligomer of n-octyl-2-cyanoacrylate (OCA), iso oligomers of -octyl-2-cyanoacrylate (IOCA), oligomers of butyl-lactoyl-2-cyanoacrylate (BLCA), and ACA oligomers of any combination or mixture thereof.

In a separate aspect, the present disclosure encompasses the use of allyl cyanoacrylates and/or aryl cyanoacrylates in the methods and products as described herein. Accordingly, in one aspect, to prepare ready-to-use poly(allyl cyanoacrylate)-based nano/microfibers instead of poly(alkyl cyanoacrylate)-based nano/microfibers, the methods described herein each allyl cyanoacrylate and poly(allyl cyanoacrylate) polymers. The method steps of such an allyl cyanoacrylate-based method are similar to the method steps of the ready-to-use poly(alkyl cyanoacrylate) based nano/microfiber manufacturing method described herein. Accordingly, the present disclosure provides allyl cyanoacrylate-related and poly( allyl cyanoacrylate) polymer-related aspects and embodiments. Such allyl cyanoacrylate-related and poly(allyl cyanoacrylate) polymer-related aspects and embodiments are specifically directed to the corresponding preparation of ready-to-use poly(allyl cyanoacrylate) based nano/microfibers. produced from, or otherwise derived from, the poly(allyl cyanoacrylate)-based nano/microfibers obtainable by the process, as well as the poly(allyl cyanoacrylate)-based nano/microfibers thus obtained; , the products and uses on which it may be based.

Likewise, in one aspect, to prepare ready-to-use poly(aryl cyanoacrylate)-based nano/microfibers instead of poly(alkyl cyanoacrylate)-based nano/microfibers, the methods described herein each cyanoacrylate and poly(aryl cyanoacrylate) polymers. The method steps of such an aryl cyanoacrylate-based method are similar to the method steps of the ready-to-use poly(alkyl cyanoacrylate) based nano/microfiber manufacturing method described herein. Accordingly, the present disclosure provides for aryl cyanoacrylate-related and poly( aryl cyanoacrylate) polymer-related aspects and embodiments. Such aryl cyanoacrylate-related and poly(aryl cyanoacrylate) polymer-related aspects and embodiments are specifically directed to the corresponding preparation of ready-to-use poly(aryl cyanoacrylate) based nano/microfibers. produced from, or otherwise derived from, the poly(aryl cyanoacrylate)-based nano/microfibers obtainable by the process, as well as the poly(aryl cyanoacrylate)-based nano/microfibers thus obtained; , the products and uses on which it may be based.

As used herein, the terms "alkyl cyanoacrylate" and "cyanoacrylate" may be used interchangeably. As described in greater detail elsewhere herein, the alkyl substituent of an “alkyl cyanoacrylate” may be unsubstituted or may be optionally substituted. Substituents may include heteroatoms and functional groups such as hydroxy groups, substituents containing ether linkages, carboxy groups, and any combination or mixture thereof. A more preferred substituent comprising a heteroatom is exemplified by the monomer butyl-lactoyl-2-cyanoacrylate (BLCA). In a preferred embodiment, the substituents contain only carbon and hydrogen atoms.

In various embodiments, the method of making (ready-to-use) PACA-based nano/microfibers or PACA-based nano/microfiber meshes comprises in step (a) suitably one single ACA monomer species or one single ACA single PACA-homopolymers composed of oligomeric species. In other words, such a method does not involve any other PACA-homopolymer species. In such an embodiment, the PACA-based nano/microfibers or PACA-based nano/microfiber mesh produced by the method of the present invention consists of one single PACA-homopolymer, i.e. any other PACA-homopolymer. species are not included. In addition, such embodiments are characterized by a polydispersity index attributable to a single PACA-homopolymer. As described elsewhere herein, the polydispersity index encompasses the ratio of the weight-average molecular weight (M _{w ) to the number-average molecular weight (M n ) of the PACA homopolymer that is between 1 and 1.7.} In various embodiments, the polydispersity index is based on one particular number-average molecular weight (M _n ), ^{which can be any of 10 3} to 10 ^{6 Da.} In other words, in various embodiments, a single PACA homopolymer species composed of one single ACA monomer (or one single ACA oligomer) has one particular number-average, which can be any ^{of 10 3} to 10 ^{6 Da.} It has a molecular weight (M _n ). Preferably, said one specific number-average molecular weight (M _n ) may be any of 10 ⁴ to 10 ⁶ Da, preferably 10 ⁴ Da or 10 ⁶ Da. In various preferred embodiments, one particular number-average molecular weight (M _n ) is 10 ⁵ or 10 ⁶ Da. In other embodiments, the polydispersity index is based on different number-average molecular weights (M _n ), ^{which can be any of 10 3} to 10 ^{6 Da.} In other words, in various embodiments, a single ACA monomers one type of single consisting of (or a single ACA oligomer of one kind) PACA different numbers that can be of any homopolymer species is 10 ³ to 10 ⁶ Da - average molecular weight ( M _n ). Preferably, said different number-average molecular weight (M _n ) may be any of 10 ⁴ to 10 ⁶ Da, preferably 10 ⁴ Da or 10 ⁶ Da. In various preferred embodiments, the different number-average molecular weights (M _n ) are 10 ⁵ and 10 ⁶ Da. In various other embodiments, a single PACA homopolymer species composed of one single ACA monomer (or one single ACA oligomeric species) has different proportions of different number-average molecular weights (M _n ), in which case a single PACA The polydispersity index of the homopolymer species is between 1 and 1.7.

In various other embodiments, the method of making (ready-to-use) PACA-based nano/microfibers or PACA-based nano/microfiber meshes comprises in step (a) at least two different ACA monomer species or different ACA oligomeric species. species PACA homopolymer species. In such embodiments, the PACA-based nano/microfibers or PACA-based nano/microfiber meshes produced by the methods of the present invention are composed of at least two PACAs alone composed of different ACA monomer species (or different ACA oligomeric species). It is composed of polymer species. In addition, such embodiments are characterized by a polydispersity index attributable to at least two PACA-homopolymer species. The polydispersity index encompasses the ratio of the weight-average molecular weight (M _{w ) to the number-average molecular weight (M n} ) of at least two PACA homopolymer species that is between 1 and 1.7. The polydispersity index is based on _{the different number-average molecular weights (M n} ) of said at least two PACA homopolymer species, which may be any ^{of 10 3} to 10 ^{6 Da.} Preferably, the number-average molecular weight (M _n ) may be any of 10 ⁴ to 10 ⁶ Da, preferably 10 ⁴ Da or 10 ⁶ Da. In various preferred embodiments, the different number-average molecular weights (M _n ) are 10 ⁵ and 10 ⁶ Da. On the other hand also encompassed are embodiments wherein the polydispersity index of at least two PACA-homopolymer species is based on one particular number-average molecular weight (M _n ), ^{which may be any of 10 3} to 10 ^{6 Da.} am. In other words, in such an embodiment, both at least two PACA homopolymer species composed of different ACA monomers (or different ACA oligomers) have the same specific number-average molecular weight, which can be any ^{of 10 3} to 10 ^{6 Da.} (M _n ). Preferably, the number-average molecular weight (M _n ) may be any of 10 ⁴ to 10 ⁶ Da, preferably 10 ⁴ Da or 10 ⁶ Da. In various preferred embodiments, the different number-average molecular weights (M _n ) are 10 ⁵ and 10 ⁶ Da. In various embodiments, the polydispersity index of at least two PACA homopolymer species composed of different ACA monomer species (or different ACA oligomeric species) is based on different proportions of different number-average molecular weights (M _n ), In this case the polydispersity index of the at least two PACA homopolymer species is between 1 and 1.7. In various embodiments, the polydispersity index, particularly the polydispersity index of 1 to 1.7, is a weight-average molecular weight (M _w ) relative to _{the number-average molecular weight (M n ) of a PACA polymer that is a PACA homopolymer and/or a PACA copolymer.} refers to the rain of

In a preferred embodiment of the aforementioned method for producing (ready-to-use) PACA-based nano/microfibers or PACA-based nano/microfiber meshes comprising at least two PACA homopolymer species in step (a), the method comprises in step (a) any of 2, 3, 4 and 5 PACA homopolymer species consisting of different ACA monomer species or different ACA oligomeric species. More preferably, the method comprises in step (a) two or three PACA homopolymer species composed of different ACA monomer species or different ACA oligomeric species.

In various embodiments, the method for making (ready-to-use) PACA-based nano/microfibers or PACA-based nano/microfiber meshes comprises in step (a) at least two different ACA monomer species or at least two single PACA copolymers composed of different ACA oligomeric species. In other words, such a method does not involve any other PACA copolymer species. In such an embodiment, the PACA-based nano/microfiber or PACA-based nano/microfiber mesh produced by the method of the present invention consists of one single PACA copolymer, i.e. it does not contain any other PACA copolymer species. does not In a preferred embodiment, said single PACA copolymer consists of any of 2, 3, 4 and/or 5 different ACA monomer species, or any of 2, 3, 4 and/or 5 different ACA oligomeric species. is composed of More preferably, said single PACA copolymer consists of two or three different ACA monomer species, or consists of two or three different ACA oligomeric species.

In addition, such embodiments are characterized by a polydispersity index attributable to a single PACA copolymer. As described elsewhere herein, the polydispersity index encompasses the ratio of the weight-average molecular weight (M _{w ) to the number-average molecular weight (M n ) of the PACA copolymer that is between 1 and 1.7.} In various embodiments, the polydispersity index is based on one particular number-average molecular weight (M _n ), ^{which can be any of 10 3} to 10 ^{6 Da.} In other words, in various embodiments, a single PACA copolymer species composed of at least two different ACA monomer species or at least two different ACA oligomeric species has one specified number, which may be any ^{of 10 3} to 10 ^{6 Da.} - has an average molecular weight (M _n ). Preferably, said one specific number-average molecular weight (M _n ) may be any of 10 ⁴ to 10 ⁶ Da, preferably 10 ⁴ Da or 10 ⁶ Da. In various preferred embodiments, one particular number-average molecular weight (M _n ) is 10 ⁵ or 10 ⁶ Da. In other embodiments, the polydispersity index is based on different number-average molecular weights (M _n ), ^{which can be any of 10 3} to 10 ^{6 Da.} In other words, in various embodiments, a single PACA copolymer species composed of at least two different ACA monomer species or at least two different ACA oligomeric species has different number-averages, which can be any ^{of 10 3} to 10 ^{6 Da.} It has a molecular weight (M _n ). Preferably, said different number-average molecular weight (M _n ) may be any of 10 ⁴ to 10 ⁶ Da, preferably 10 ⁴ Da or 10 ⁶ Da. In various preferred embodiments, the different number-average molecular weights (M _n ) are 10 ⁵ and 10 ⁶ Da. In various other embodiments, a single PACA copolymer species composed of at least two different ACA monomer species or at least two different ACA oligomeric species has different proportions of different number-average molecular weights (M _n ), in which case single The polydispersity index of the PACA copolymer species ranges from 1 to 1.7.

In various other embodiments, the method of making (ready-to-use) PACA-based nano/microfibers or PACA-based nano/microfiber meshes comprises in step (a) at least two different ACA monomer species or different ACA oligomeric species. species of PACA copolymers. In such embodiments, the PACA-based nano/microfibers or PACA-based nano/microfiber meshes produced by the methods of the present invention comprise at least two PACA polymers composed of different ACA monomer species (or different ACA oligomeric species). composed of synthetic species. In addition, such embodiments are characterized by a polydispersity index attributable to at least two PACA copolymer species. The polydispersity index encompasses the ratio of the weight-average molecular weight (M _{w ) to the number-average molecular weight (M n} ) of at least two PACA copolymer species that is between 1 and 1.7. The polydispersity index is _{based on the different number-average molecular weights (M n} ) of said at least two PACA copolymer species, which can be any ^{of 10 3} to 10 ^{6 Da.} Preferably, the number-average molecular weight (M _n ) may be any of 10 ⁴ to 10 ⁶ Da, preferably 10 ⁴ Da or 10 ⁶ Da. In various preferred embodiments, the different number-average molecular weights (M _n ) are 10 ⁵ and 10 ⁶ Da. Also encompassed on the other hand are embodiments in which the polydispersity index of at least two PACA copolymer species is based on one particular number-average molecular weight (M _n ), ^{which may be any of 10 3} to 10 ^{6 Da.} . In other words, in such an embodiment, both at least two species of PACA copolymers composed of different ACA monomer species or different ACA oligomeric species have the same specific number-average molecular weight, which may be any ^{of 10 3} to 10 ^{6 Da.} (M _n ). Preferably, the number-average molecular weight (M _n ) may be any of 10 ⁴ to 10 ⁶ Da, preferably 10 ⁴ Da or 10 ⁶ Da. In various preferred embodiments, the different number-average molecular weights (M _n ) are 10 ⁵ and 10 ⁶ Da. In various embodiments, the polydispersity index of at least two PACA copolymer species composed of different ACA monomer species (or different ACA oligomeric species) is based on different proportions of different number-average molecular weights (M _n ), In this case the polydispersity index of the at least two PACA copolymer species is between 1 and 1.7. In a preferred embodiment of the aforementioned method for producing (ready-to-use) PACA-based nano/microfibers or PACA-based nano/microfiber meshes comprising at least two PACA copolymer species in step (a), the method comprises in step (a) any of 2, 3, 4 and 5 PACA copolymer species consisting of different ACA monomer species or different ACA oligomeric species. More preferably, the method comprises in step (a) two or three PACA copolymer species composed of different ACA monomer species or different ACA oligomeric species.

As described herein, an ACA oligomer is any of 2 to 10 ACA monomer units, including dimers, trimers, tetramers, pentamers, hexamers, heptomers, octamers, tetramers and de-mers as appropriate. can be configured.

PACA polymers as used in the present invention, preferably high molecular weight PACA (homo)polymers (eg PBCA) can be prepared by anionic bulk or mass polymerization. High molecular weight PACA (homo)polymers may be characterized by a molecular weight ranging ^{from 10 4} to 10 ^{6 Da.} Preferably, the molecular weight represents a number-average molecular weight. DMPT may also be used as an initiator in anionic polymerizations, especially in anionic bulk or bulk polymerizations. In various embodiments of the present invention, the anionic polymerization as disclosed herein is carried out in the presence of an initiator or catalyst. In various embodiments, the initiator or catalyst is an anionic initiator or anionic catalyst. In various embodiments, the initiator or catalyst is a tertiary amine, preferably an aromatic amine. A-toluidine (DMPT) - In various preferred embodiments, the initiator or catalyst is N, N '- dimethyl - p. In various embodiments, the initiator is dissolved in a solvent (1/1000, v/v). Preferably, the solvent is acetone. Anionic (bulk or bulk) polymerization can be carried out by first adding the ACA to a reaction chamber (such as a flask, more specifically a 3-neck round-low flask), followed by the addition of an initiator. Preferably, the polymerization reaction is carried out in a low temperature reaction system (eg, the polymerization reaction may be performed on a low temperature water bath). At the end of the polymerization, the solid formed may be dissolved, precipitated in distilled water, filtered, washed first with water, then with methanol, and dried in vacuo. As Table 1 below, the identification of PACA using number-average molecular weight values and molar combinations between ACA and initiator can be illustrated.

Bulk polymerization or bulk polymerization can be carried out by adding a soluble (radical) initiator to the pure monomers in liquid state. The initiator must be dissolved in the monomer. The reaction is initiated by heating or exposure to radiation. As the reaction progresses, the mixture becomes more viscous. The reaction is exothermic and yields a wide range of molecular masses. In various embodiments of the present invention, the anionic polymerization of ACA is carried out in bulk (ie with pure ACA monomer in liquid state). In various other embodiments of the present invention, the anionic polymerization is carried out using ACA prepared from ACA oligomers resulting from the Neufenagel condensation of an alkyl cyanoacetate with formaldehyde followed by pyrolysis of the formed oligomeric mixture. PACAs can be obtained via direct Neubenagel condensation between the corresponding alkyl cyanoacetates and formaldehyde, or by anionic polymerization of their monomers. 7 shows GPC results of electro-spun mesh of A) powder of PACA, B) powder of PACA-co-CS and C) PACA/PACA-co-CS.

Polymerization of ACA using an anionic initiator, preferably a tertiary amine, more preferably DMPT, has been successfully carried out over a wide range of polymer/initiator ratios. In various embodiments of the present invention, the ACA and initiator/catalyst can be applied to the anionic polymerization reaction in a specific ratio. For example, in various embodiments, the ratio of ACA:initiator/catalyst ^{may be about 1/10 3} :1. In various other embodiments, the ratio of ACA:initiator/catalyst ^{can be about 2/10 2} :1. This ratio allows the preparation of PACAs with number-average molecular weights between ^{10 4} and 10 ^{6 Da.} The number-average molecular weight can be defined as the total weight (mass) of the polymer divided by the total number of molecules (the arithmetic mean of the molar mass distribution). The weight-average molecular weight relates to the sum of all molecular weights (the sum of the products of the molar masses of each fraction) times its weight fraction. The polydispersity index provides a measure of the molecular weight distribution within a sample. It has a value greater than 1; It is equal to 1 only if all molecules have the same weight (ie they are monodisperse), and the greater the spread of the molecular weight, the further it is. In the process of the present invention, well-defined polymeric materials are used rather than polymer mixtures or copolymer mixtures comprising widely different molecular weights.

In various preferred embodiments of the present invention, the ACA is BCA, the initiator is a tertiary amine, preferably the initiator is DMPT.

In various embodiments, the electrospinning parameters according to the present disclosure may be any one of the following: voltage = 5-30 kV, preferably 5-22 kV; flow = 0.025 to 2.000 mL/min, preferably 0.025-0.125 mL/min; needle tip diameter = 0.3-1 mm; needle tip-collector distance = 5-30 cm, preferably 5-15 cm; polymer concentration = 0.5-50 % w/w; and/or solvent mixture = acetone:ethanol 1:0 to 4:1 (v/v). During the electrospinning step, the polymer solution jet (or polymer melt) is charged by an electrical potential that breaks the surface tension of the solution droplet at the needle tip end due to the repulsion of the same charge. This induces an opposite power or charged jet transfer to the ground-connected collector. In its most basic form, an electrospinning machine consists of a high voltage power source, an infusion system pump, a syringe equipped with a needle and a collector. The shape of the droplet at the moment of breakage is referred to as a “Taylor cone” (conical surface of the solution at the needle tip). The charged jet is then dried on its journey to the collector and stretched as the solvent evaporates to form very thin fibers in the nanometer to micrometer range. 1 shows a typical electrospinning apparatus: A) a high voltage power supply, B) an injection system pump and C) a collector connected to ground. Figure 2 shows an infusion system pump with a syringe associated with a needle and a ground connected collector; In general, typical collectors suitable for the method according to the present invention include, but are not limited to, copper mesh, copper ring, aluminum foil, paper and stainless steel rectangle. 3 shows an electro-spun mesh of PACA substrate collected on a copper mesh used as a collector. Figure 4 shows an electro-spun mesh of PACA substrate collected on different collectors comprising A) copper rings, B) aluminum foil and C) paper.

Surprisingly, a solution comprising (i) a PACA homopolymer and/or a PACA copolymer and (ii) a non-PACA polymer (which may be a non-PACA homopolymer and/or a non-PACA copolymer) is a ready-to-use PACA It has been found that it can be successfully applied to electrospinning to produce nano/microfiber-based and/or PACA-based nano/microfiber meshes. Accordingly, in various embodiments, the method for producing (ready-to-use) PACA-based nano/microfibers and/or PACA-based nano/microfiber meshes comprises in step (b) prior to electrospinning in step (b). at least one non-PACA polymer (non-PACA homopolymer and/or non-PACA homopolymer and/or non-PACA polymer) into the solution obtained, ie the solution obtained by dissolving the PACA homopolymer and/or the PACA copolymer of step (a) in a solvent PACA copolymer). In various embodiments, the non-PACA polymer (which may be a non-PACA homopolymer and/or a non-PACA copolymer) is poly(lactic acid-co-glycolic acid) (PLGA), poly(ε-caprolactone) ( PCL), poly(ethylene glycol) (PEG), poly(lactic acid) (PLA), chitosan (CH), hyaluronic acid (HA), dextran (Dex), and any combination or mixture thereof, However, the present invention is not limited thereto. In various embodiments, the non-PACA copolymer is poly(lactic acid-co-glycolic acid) (PLGA) or poly(lactic acid) (PLA). In various preferred embodiments, the non-PACA homopolymer is poly(lactic acid) (PLA). In various other preferred embodiments, the non-PACA homopolymer is chitosan or PLGA.

In various other embodiments, the non-PACA polymer (which may be a non-PACA homopolymer and/or a non-PACA copolymer) is a bioadhesive non-PACA polymer (a bioadhesive non-PACA homopolymer and/or a non-PACA copolymer). or bioadhesive non-PACA copolymers). As used herein, bioadhesion refers to the ability of certain synthetic and biological macromolecules to adhere to biological tissues. Bioadhesion may depend, in part, on the properties of the polymer, biological tissue and the surrounding environment. It has been found that several factors contribute to the bioadhesive ability of polymers: the presence of functional groups (-OH, -COOH) capable of forming hydrogen bridges, sufficient elasticity for the polymer chains to interpenetrate into the mucus layer, and high molecular weight. Accordingly, in various embodiments, the bioadhesive non-PACA polymer is characterized by the presence of one or more functional groups capable of forming hydrogen bridges, such as -OH, -COOH. Bioadhesive systems are being used in dentistry, orthopedics, ophthalmology and surgical applications.

The amount of non-PACA polymer (which may be non-PACA homopolymer and/or non-PACA copolymer) added to a solution comprising PACA homopolymer and/or PACA copolymer can vary widely, typically ( from about 0.1% to about 75% by weight of a solution comprising i) a PACA homopolymer and/or a PACA copolymer and (ii) a non-PACA polymer (which may be a non-PACA homopolymer and/or a non-PACA copolymer). % by weight. In some embodiments, the concentration of the non-PACA polymer (which may be a non-PACA homopolymer and/or a non-PACA copolymer) is in the range of about 0.5-5% by weight; In some embodiments, the concentration of the non-PACA polymer (which may be a non-PACA homopolymer and/or a non-PACA copolymer) is in the range of about 0.7-4% by weight; In some embodiments, the concentration of the non-PACA polymer (which may be a non-PACA homopolymer and/or a non-PACA copolymer) is in the range of about 1-3% by weight. In various other embodiments, the concentration of the non-PACA polymer (which may be a non-PACA homopolymer and/or a non-PACA copolymer) is in the range of about 25-75 weight percent. Preferably when the non-PACA homopolymer is PLGA, more preferably when the non-PACA homopolymer is PLGA and the PACA is PBCA, the non-PACA polymer (non-PACA homopolymer and/or non-PACA copolymer) may be in the range of about 25-75% by weight.

In some embodiments, the concentration of the non-PACA polymer (which may be a non-PACA homopolymer and/or a non-PACA copolymer) is in the range of about 0.7-4% by weight; In some embodiments, the concentration of the non-PACA polymer (which may be a non-PACA homopolymer and/or a non-PACA copolymer) is in the range of about 1-3% by weight.

As described elsewhere herein, the poly(alkyl cyanoacrylate) copolymer in step (a) of the method described herein comprises (i) at least two different ACA monomer species, (ii) at least at least two alkyl cyanoacrylate oligomers comprising two different ACA monomer species, or (iii) at least one poly(alkyl cyanoacrylate) (PACA) polymer and at least one non-PACA polymer. may include. Accordingly, with respect to (iii) mentioned above, the PACA copolymer may be obtainable, or obtainable, or produced by ionic polymerization of one or more ACA monomers with one or more non-PACA polymers. can be Such non-PACA polymers are capable of catalyzing or initiating the ionic polymerization of ACA monomers due to the ionic groups of the non-PACA polymers, ie, the ionic groups act as initiators of the anionic polymerization of the ACA monomers. In various embodiments, the aforementioned ionic polymerization is anionic polymerization, or cationic polymerization. Preferably, the ionic polymerization is anionic polymerization.

In various embodiments of any of the methods disclosed herein, the non-PACA polymer when used in the present invention is capable of initiating anionic polymerization of an ACA monomer. Non-PACA polymers when used in the present invention, particularly non-PACA polymers capable of initiating anionic polymerization of ACA monomers, may have anionic groups (i.e. may be anionic non-PACA polymers), or may have cationic groups may be (ie may be cationic non-PACA polymers), or may be neutral non-PACA polymers. In various preferred embodiments, the non-PACA polymer as used in the present invention may be an organic non-PACA polymer, preferably an organic anionic or organic cationic non-PACA polymer, ie each may have an organic anionic group or , or an organic cationic group. The present disclosure also encompasses organic neutral non-PACA polymers. As described herein, anionic groups can be considered groups that retain an overall negative charge. Likewise, a cationic group can be considered a group that retains an overall positive charge. A neutral non-PACA polymer can also be considered a non-PACA polymer with an overall neutral charge. Also, any anionic non-PACA polymer as described herein may be considered an anionic non-PACA polymer having an overall negative charge. Likewise, any cationic non-PACA polymer as described herein may be considered a cationic non-PACA polymer having an overall positive charge.

In a preferred embodiment, the (organic) anionic groups of the anionic non-PACA polymers described herein may be amino groups, or hydroxyl groups, or carboxyl groups. In various embodiments, non-PACA polymers as described herein can be polysaccharides, glycosaminoglycans, or polyethers. In a preferred embodiment, the non-PACA polymer as described herein is a polysaccharide. The polysaccharides may be considered neutral, cationic or anionic polysaccharides. Likewise, the glycosaminoglycans and polyethers may be considered neutral, cationic or anionic glycosaminoglycans or polyethers. In a more preferred embodiment, the non-PACA polymer as described herein may be any of polyethylene glycol (PEG), chitosan, dextran and hyaluronic acid. In a particularly preferred embodiment, the non-PACA polymer is chitosan. In a preferred embodiment, any of the non-PACA polymers as described herein are biodegradable non-PACA polymers. The term "biodegradable non-PACA polymer" may be considered a non-PACA polymer that is degradable by biological activity. More specifically, the biological activity may be enzymatic hydrolysis, or degradation by microorganisms, particularly bacteria. Degradation by microorganisms, particularly bacteria, may include enzymatic hydrolysis.

As further described herein, any of the PACA (homo)polymers disclosed herein may also be considered biodegradable PACA (homo)polymers. The term "biodegradable PACA polymer" may be considered a PACA (homo)polymer that is degradable by biological activity. More specifically, the biological activity may be enzymatic hydrolysis, or degradation by microorganisms, particularly bacteria. Degradation by microorganisms, particularly bacteria, may include enzymatic hydrolysis.

Chitosan is produced by the deacetylation of chitin, a structural component of the crustacean exoskeleton and fungal cell walls. Chitosan is a polymer composed of randomly distributed β-(1,4)-linked D-glucosamine (deacetylated units) and N -acetyl-D-glucosamine (acetylated units). Chitin is a useful biocompatible material that can be used in medical applications including, but not limited to, tissue regeneration and tissue engineering. Chitosan materials contemplated for use herein may also be characterized by the percent deacetylation of that chitosan, which in some embodiments may be up to 100%; In some embodiments, the chitosan has a percent deacetylation of from about 65 to about 100%; In some embodiments, the chitosan has a percent deacetylation of from about 70 to about 100%; In some embodiments, the chitosan has a percent deacetylation of from about 75 to about 100%; In some embodiments, the chitosan has a percent deacetylation of from about 80 to about 100%. In some embodiments, chitosan contemplated for use herein may be from any source of animal or microbial origin; In some embodiments, the chitosan contemplated for use herein is of crustacean origin. As will be appreciated by those of ordinary skill in the art, the amount of chitosan present in the formulations contemplated herein can vary widely, typically (i) PACA homopolymers and/or PACA copolymers, respectively, and (ii) chitosan, or in the range of from about 0.1% to about 30% by weight based on the weight of a solution comprising (i) a PACA (homo)polymer and (ii) a non-PACA polymer. In some embodiments, the chitosan concentration is in the range of about 0.5-5% by weight; In some embodiments, the chitosan concentration is in the range of about 0.7-4% by weight; In some embodiments, the chitosan concentration is in the range of about 1-3% by weight. In various preferred embodiments of the present disclosure, the weight ratio of chitosan:PACA is about 3-10 ^-3 :1, for example, the chitosan weight is 0.3%. Accordingly, in various preferred embodiments, the chitosan concentration is about 0.3% by weight. In the electrospun mesh shown on the right of Figure 6E, the weight ratio between PACA and PACA-co-chitosan is 50:50, ie the weight of chitosan is 0.15% of the total PACA polymer. Accordingly, in various preferred embodiments, the chitosan concentration is about 0.15% by weight. In various other preferred embodiments, the chitosan concentration ranges from about 0.15% to about 0.3% by weight.

Chitosan materials contemplated for use herein may also be characterized with respect to one or more of the following: a moisture content of <15%; Ash content <1%, turbidity <50 NTU, solubility >99%, and/or <3 ppm of heavy metals (ie As, Cd, Pb, Hg).

Chitosan-containing ready-to-use PACA-based nano/microfibers and/or PACA-based nano/microfiber meshes contemplated herein are for targeting, for example, wounds, aches and pains, skin inflammation due to insect bites, bedsores, ulcers, etc. It can be used in a variety of ways for the treatment of In some embodiments of the present invention, the chitosan-containing ready-to-use PACA-based nano/microfibers and/or PACA-based nano/microfiber meshes contemplated herein are for use, such as cosmetic and/or personal care applications for It may be suitable: (i) coating and protection as a film-forming component to aid in moisture control; (ii) providing antimicrobial activity as a natural preservative; and (iii) acting as a carrier for other active ingredients for slow release due to ionic binding to the compound and degradation of chitosan by natural skin enzymes.

As described herein, the present disclosure provides a PACA-based nano/microfiber and/or a PACA-based nano/microfiber mesh obtainable by, obtained by, or produced by any of the methods of the present invention. encompasses Because of the steps and materials (i.e. ACA monomers, and/or ACA oligomers, and/or PACA homopolymers, and/or PACA copolymers) used in the methods provided by the present invention, the inventive PACA-based nano/microfibers and Because of the structural features imparted to/or PACA-based nano/microfiber meshes, such PACA-based nano/microfibers and/or PACA-based nano/microfiber meshes are novel in the art. _{In particular, the weight-to-number-average molecular weight (M n} ) of poly(alkyl cyanoacrylate) homopolymers and/or poly(alkyl cyanoacrylate) copolymers, which may have a (polydispersity) ratio of 1 to 1.7 - The ratio of average molecular weight (M _w ) provides novel characteristics to the novel PACA-based nano/microfibers and/or PACA-based nano/microfiber meshes provided by the present invention. As described herein, the PACA-based nano/microfibers and/or the PACA-based nano/microfiber mesh provided by the present invention are the PACA-based nano/microfibers and/or PACA-based nanofibers referred to in the methods of the present disclosure. /microfiber mesh and/or any of the structural features described in relation to ACA monomers and ACA oligomers.

In some aspects, the present disclosure provides an oral cavity comprising orally applying (or administering) any of the chitosan-containing ready-to-use PACA-based nano/microfibers disclosed herein to a soft or hard tissue of a subject's mouth. A method of promoting health (eg, promoting gingival health) is provided. In a similar embodiment, the present disclosure provides any of the chitosan-containing ready-to-use PACA-based nano/microfibers disclosed herein for use in promoting oral health (such as promoting gingival health). to provide. In another similar embodiment, the present disclosure provides a medical device or medicament for promoting oral health (such as promoting gingival health) of any of the chitosan-containing ready-to-use PACA-based nano/microfibers disclosed herein. A use for manufacturing is provided. The present disclosure provides for the treatment of oral mucosa and/or dental support tissue disorders, such as implant-, comprising orally applying or administering to a subject any of the chitosan-containing ready-to-use PACA-based nano/microfibers disclosed herein. A method of treating peri-implantitis is provided.

As already described elsewhere herein, the weight-average molecular weight relative to _{the number-average molecular weight (M n} ) of the poly(alkyl cyanoacrylate) homopolymer and/or the poly(alkyl cyanoacrylate) copolymer. The ratio of (M _w ) is from 1 to 1.7. Accordingly, such ratios are considered to describe the polydispersity index of the poly(alkyl cyanoacrylate) homopolymers and/or poly(alkyl cyanoacrylate) copolymers. Preferably, said ratio of weight-average molecular weight (M _w ) to number-average molecular weight (M _n ) is between 1.1 and 1.5. More preferably, the ratio of the weight-average molecular weight (M _w ) to the number-average molecular weight (M _n ) is from 1.2 to 1.4 or from 1.4 to 1.7. In another preferred embodiment, the ratio of weight-average molecular weight (M _w ) to number-average molecular weight (M _n ) is from 1.5 to 1.7. Molecular weight can be determined using gel permeation chromatography (GPC). The measurement can be based on polystyrene or PMMA as standard using either DMF, acetone or THF as solvent. Measurements can be carried out at ambient temperature and pressure, such as 25° C. and 1 atmosphere.

The present disclosure provides PACA-based nano/microfibers or PACA-based nano/microfiber meshes made or obtainable by any of the methods described herein. Advantageously, the PACA-based nano/microfibers or PACA-based nano/microfiber meshes prepared or obtainable by any of the methods described herein include ready-to-use PACA-based nano/microfibers or ready-to-use PACA-based nano/microfibers. It can be considered a microfiber mesh. In various aspects, the PACA-based nanofibers of the present invention can have (average) diameters in the range of about 1 nm to about 999 nm, including (average) diameters in the range of between 0.01 nm and 1 nm. In various embodiments, PACA-based nanofibers of the present invention have (average) diameters in the range of from about 100 nm to about 900 nm. In various other embodiments, the PACA-based nanofibers of the present invention have a (average) diameter in the range from about 200 nm to about 800 nm. In another embodiment, the PACA-based nanofibers of the present invention have a (average) diameter in the range from about 300 nm to about 700 nm. In another embodiment, the PACA-based nanofibers of the present invention have a (average) diameter in the range of about 400 nm to about 600 nm.

In various other embodiments, the PACA-based nanofibers of the present invention have a (average) diameter in the range of about 600 nm to about 990 nm. In another embodiment, the PACA-based nanofibers of the present invention are ≤ about 600 nm, preferably in the range of about 200 nm to about 600 nm, or in the range of about 100 nm to about 600 nm, more preferably in the range of about 200 nm to about It has a (average) diameter in the range of 500 nm, or in the range of about 100 nm to about 500 nm.

In another preferred embodiment, the PACA-based nano/microfibers of the present invention have a diameter of about 1,000±300 nm, more preferably about 970±280 nm, or about 800±300 nm, more preferably about 780±300 nm ( average) diameter. In various other preferred embodiments, the PACA-based nano/microfibers of the present invention have a thickness of about 1,000 ± 200 nm, more preferably about 1,000 ± 200 nm, or about 800 ± 200 nm, more preferably about 780 ± 200 nm. (average) diameter.

Likewise, the PACA-based microfibers of the present invention may have an average diameter in the micrometer size range. In various aspects, the PACA-based microfibers of the present invention can have (average) diameters in the range of about 1 μm to about 10 μm, including (average) diameters in the range between 0.01 μm and 1 μm. In various embodiments, the PACA-based microfibers of the present invention range from about 1 μm to about 5 μm, preferably from about 1 μm to about 4 μm, more preferably from about 1 μm to about 3 μm, even more preferably from about 1 μm to about 3 μm. preferably have a (average) diameter in the range of about 1 μm to about 2 μm. In various embodiments, the PACA-based microfibers of the present invention may have a (average) diameter in the range of about 2 μm to about 5 μm, preferably in the range of about 2 μm to about 4 μm. In another embodiment, the PACA-based microfibers of the present invention may have a (average) diameter in the range of about 3 μm to about 5 μm, preferably in the range of about 3 μm to about 4 μm.

In various embodiments, the average fiber diameter of the nano/microfibers of the PACA-based nano/microfibers or PACA-based nano/microfiber mesh can be between 1 nm and 500 μm. Preferably, the average fiber diameter of the nano/microfibers of the PACA-based nano/microfibers or the PACA-based nano/microfiber mesh may be from 10 nm to 50 μm. More preferably, the average fiber diameter of the nano/microfibers of the PACA-based nano/microfibers or the PACA-based nano/microfiber mesh may be 100 nm to 5 μm.

In various embodiments, the novel PACA-based nano/microfibers and/or PACA-based nano/microfiber meshes of the present invention may be chemically and/or physically modified for biomedical applications, preferably in this case the present invention. The surfaces of the novel PACA-based nano/microfibers and PACA-based nano/microfiber meshes are modified by encapsulation and/or immobilization of therapeutic agents and/or biological materials. Specifically, in various embodiments, the novel PACA-based nano/microfibers and/or PACA-based nano/microfiber meshes of the present invention may contain therapeutic agents, antibiotics, antiviral agents, chemotherapeutic agents, analgesic and analgesic combinations, anti-inflammatory agents, vitamins, sedatives, Radiopharmaceutical, metal particle, metal alloy particle, metal oxide particle, hydroxyapatite, sodium alginate, dye, cell, protein, peptide, nucleic acid, nucleic acid analog, nucleotide or oligonucleotide, peptide nucleic acid, aptamer, antibody, or its at least one of fragments or portions, antigens or epitopes, hormones, hormone antagonists, growth factors or recombinant growth factors, and fragments and variants thereof, cell adhesion mediators, cytokines, enzymes, and/or any combination or mixture thereof; They may be fixed, or they may be enclosed, or they may be dotted with them.

The therapeutic agent may be, for example, dissolved in a solution to be electro-spun, alternatively co-electrospun, or used for coaxial electro-spinning into a fiber core. It can be homogeneously distributed over the body of the nano/microfiber(s) and/or mesh(s) using simultaneous electrospinning of a PACA solution and electrospraying of a therapeutic agent solution, or PACA(s) and The PACA-based nano/microfiber(s) and/or microparticles may be loaded into the PACA-based nano/microfiber(s) and/or microparticles homogeneously distributed into the mesh by electrospinning of one emulsion comprising therapeutic agent-loaded microparticles, or by adsorption or chemical bonding to the mesh. may be coated on the surface of (s). The method and amount of incorporation of a therapeutic agent into the nano/microfiber(s) and/or mesh(s) will depend upon the nature of the agent and the biomedical application to be performed.

Meshes according to the present invention are suitable for a variety of applications including biomedical products for wound dressing/healing, tissue regeneration/engineering, membranes for cell separation, organ protection, implant coating, controlled drug delivery and in vitro cell culture . Preferred applications include dental applications, particularly applications in dental surgery.

In various embodiments, the novel PACA-based nano/microfibers and/or PACA-based nano/microfiber meshes of the present invention may comprise live cells seeded on/on the mesh or nano/microfibers. The cell may be any type of animal cell, such as a human cell, or even a cell from a subject to be treated. In various embodiments, the novel PACA-based nano/microfibers and/or PACA-based nano/microfiber meshes of the present invention may comprise a cell growth medium.

The inclusion of any active agent or combination of active agents makes it possible to use the mesh for a number of biomedical purposes. The ready-to-use nano/microfiber mesh of the present disclosure is suitable for delivery of load molecules such as pharmaceutically or cosmetically active agents and nutritional supplements (also commonly referred to herein as “active agents”). In various embodiments, the active agent is soluble in the same solvent as the PACA or PACA-based nano/microfiber mesh. This allows, but is not required, a fiber or mesh in which the active agent is completely homogeneous in the PACA matrix, using electro-spinning to encapsulate any type of active agent (hydrophobic and hydrophilic), e.g. polymers and active agents. Other strategies exist to prepare a solvent mixture capable of dissolving all of the agents, or to prepare an emulsion consisting of (1) a polymer that will form fibers and (2) polymer particles that charge the active agent; This is because it is also possible to configure coaxial or triaxial electrospinning. Alternatively, the active agent can be physically or chemically immobilized on the prepared electro-spun mesh.

The ready-to-use nano/microfiber mesh of the present disclosure protects the agent(s) from premature degradation and enables sustained release thereof, thereby increasing the bioavailability of the immobilized or encapsulated active agent(s) and can increase efficacy. The ready-to-use nano/microfiber meshes of the present invention can advantageously exhibit high loadings of any load molecules, (therapeutic/active) agents and/or biological materials. Accordingly, the ready-to-use nano/microfiber mesh of the present invention contains at least 5 wt-%, about 5- 10 wt-%, about 10-15 wt-%, about 15-20 wt-%, about 20-25 wt-%, about 25-30 wt-% of any active agent(s), therapeutic agent(s) and / or biological material. In various embodiments, the ready-to-use nano/microfiber mesh of the present invention comprises greater than 30 wt-% relative to the total weight of the poly(alkyl cyanoacrylate) polymer(s) of the ready-to-use nano/microfiber mesh. may include any active agent(s), therapeutic agent(s) and/or biological material. In a preferred embodiment, the active agent is 5FU and/or the PACA is PBCA.

The present invention provides a nano/microfiber mesh or nano/microfiber network comprising the novel PACA-based nano/microfibers disclosed herein. As used herein, the terms "nano/microfiber mesh" and "nano/microfiber network" may be used interchangeably.

The present invention also relates to (i) a PACA-based nano/microfiber or PACA-based nano/microfiber mesh disclosed herein and (ii) a non-PACA polymer (which may be a non-PACA homopolymer and/or a non-PACA copolymer). ) to provide a composite material comprising Characteristics and embodiments of the above non-PACA polymers (which may be non-PACA homopolymers and/or non-PACA copolymers) are described elsewhere herein and in the composite materials provided by the present invention. is encompassed as In various preferred embodiments, the composite material comprises (i) a PACA-based nano/microfiber or PACA-based nano/microfiber mesh disclosed herein and (ii) chitosan or PLGA.

As described hereinabove, the present invention demonstrates that a mesh or mesh network of PACA nano/microfibers can be prepared by electrospinning a PACA solution. As shown in the examples and the accompanying drawings, different types of meshes were fabricated. Specifically, it has been shown that meshes, for example, can be made based on PACA alone, in particular on the basis of PBCA (poly( n -butyl cyanoacrylate)) alone.

PACAs with different molecular weights from high molecular weight (10 ⁶ Da) to low molecular weight (10 ³ Da) can be used for electrospinning. Preferably, the molecular weight represents a number-average molecular weight. In various embodiments of the present invention, the PACA polymer concentration, particularly the PBCA concentration, is preferably between 0.5 and 50 % w/w in acetone or an acetone/ethanol mixture. In various embodiments, the PACA is ^{a PACA homopolymer, particularly a PBCA homopolymer, having a high molecular weight, such as 10 5} Da or 10 ⁶ Da, the polymer concentration (PACA, particularly PBCA) being between 0.5 and 15% w/w in solution. is the range In various preferred embodiments, the solvent is acetone or an acetone/ethanol mixture. Preferably, the polymer concentration (PACA, in particular PBCA) of the high molecular weight polymer is in the range between 0.5 and 10 % w/w, wherein the solvent is preferably acetone or an acetone/ethanol mixture. More preferably, the polymer concentration (PACA, especially PBCA) of the high molecular weight polymer is in the range between 0.5 and 5 % w/w, wherein the solvent is preferably acetone or an acetone/ethanol mixture. Even more preferably, the polymer concentration (PACA, in particular PBCA) of the high molecular weight polymer ranges between 0.5 and 2 % w/w or between 0.5 and 1.5 % w/w, wherein the solvent is preferably acetone or acetone/ It is an ethanol mixture. In a particularly preferred embodiment, the high molecular weight polymer (PACA, in particular PBCA) has a concentration of about 0.5 % w/w, wherein the solvent is preferably acetone or an acetone/ethanol mixture. Such embodiments may result in the formation of PACA nanofibers (particularly PBCA nanofibers) having average fiber diameters in the nanometer and micrometer range. For example, a mesh prepared using a PACA (particularly PBCA) polymer concentration of 0.5% w/w (the solvent is preferably acetone or an acetone/ethanol mixture) may be a PACA nanofiber having an average fiber diameter of about 600 nm. (especially PBCA nanofibers). Such meshes are encompassed by the present disclosure. In addition, meshes prepared using a PACA (particularly PBCA) polymer concentration of 1.1% w/w (the solvent is preferably acetone or an acetone/ethanol mixture) have an average fiber diameter of about 1.1 μm with PACA nanofibers (particularly PBCA nanofibers). Such meshes are also encompassed by the present disclosure. Thus, the fiber diameter of the mesh increases with the concentration of the polymer solution. In various embodiments of the present invention, when aiming to produce PACA-based fibers having average fiber diameters in the nanometer range, the PACA polymer concentration is from 0.1 % w/w to about 1.0 % w/w of a polymer concentration (solvent is preferably acetone or an acetone/ethanol mixture). In various other embodiments of the present invention, when aiming to produce PACA-based fibers having an average fiber diameter in the micrometer range, the PACA polymer concentration is a polymer concentration of ≧1.1% w/w (the solvent is preferably acetone). or an acetone/ethanol mixture). The thickness of the mesh as described in this paragraph is on the order of 260 μm. Also, the Young's modulus of this mesh may be about 1.2 MPa, while the tensile strength may be about 0.08 MPa, and the failure strain may be about 15%. Thus, the mechanical properties of the mesh provided by the present invention are closer to elastic materials than rigid materials.

Surprisingly, using a very low viscosity (0.3 - 3 cP) PACA solution (particularly a PBCA solution) as described in the preceding paragraph, which is unknown to our knowledge or not possible using other polymers such as PCL, , it has been demonstrated that continuous and smooth fibers can be obtained. Accordingly, in various embodiments of the present invention, the PACA solutions (particularly PBCA solutions) disclosed herein exhibit a viscosity of less than 10 cP, preferably less than 5 cP, more preferably less than 3 cP. In various preferred embodiments of the present invention, the PACA solutions disclosed herein (particularly PBCA solutions) exhibit a viscosity in the range between 0.3 and 3 cP.

In various embodiments, a mesh based on a blend solution of a high molecular weight PACA polymer, preferably a high molecular weight PACA homopolymer (such as PBCA) and a non-PACA polymer, preferably a non-PACA homopolymer (such as chitosan), is about It can have an average fiber diameter of 1 μm and a thickness of about 400 μm, a Young's modulus of about 1 MPa, a tensile strength of about 0.07 MPa, and a strain to failure of 14%. Such a mesh may also have a minimum pore area of ^{0.1 μm 2} and a maximum pore area of ^{about 482 μm 2 .}

In various other embodiments, a mesh based on a blend solution of a high molecular weight PACA polymer, preferably a high molecular weight PACA homopolymer (such as PBCA) and a non-PACA polymer, preferably a non-PACA copolymer (such as PLGA), comprises: It can have an average fiber diameter of about 900 nm and a thickness of about 390 μm, a Young's modulus of about 1 MPa, a tensile strength of about 0.06 MPa, and a strain to failure of 20%. Such a mesh may also have a pore area in the range of ^{0.1 μm 2} to 160 μm ^{2 .}

In various other embodiments, a mesh based on a solution of a high molecular weight PACA polymer, preferably a high molecular weight PACA homopolymer (such as PBCA) and a therapeutic agent, such as 5FU, has an average fiber diameter ranging between 700 nm and 1.2 μm and about 400 It can have a thickness of μm, a Young's modulus of about 0.7 MPa, a tensile strength of about 0.1 MPa, and a strain to failure of 25%. Preferably 7, 10, 15, 20, 25 and 30% by mass of the PACA polymer, which is a high molecular weight PACA (homo)polymer, and the average fiber diameter increases with increasing drug concentration (from 700 nm to 1.2 μm). Preferably, such high molecular weight PACA (homo)polymers are dissolved in the solvent at a concentration of 1-2% w/w. Preferably, the solvent is acetone.

As above, in various embodiments of the present invention, nano/microfiber meshes as disclosed herein, preferably meshes made on the basis of high molecular weight PACA (homo)polymers, are in the micrometer range, preferably 700 It has a thickness of less than μm, more preferably less than 500 μm, even more preferably less than 400 μm, even more preferably less than 300 μm. In various embodiments, the thickness is ≤ 260 μm. In various embodiments of the present invention, the pore area of the meshes disclosed herein ranges ^{from 0.1-370 μm 2 .} In various embodiments, the average fibers of the mesh in question have a thickness in the micrometer range, preferably less than 700 μm, more preferably less than 500 μm, even more preferably less than 400 μm, even more preferably less than 300 μm. The diameter may range between 500 nm and 1.5 μm. In various other embodiments, such meshes may also have a Young's modulus in the range of about 0.5 MPa to 1.5 MPa. In another embodiment, such a mesh may also have a tensile strength in the range of about 0.05 MPa to 0.15 MPa. In various preferred embodiments, a mesh as disclosed herein, preferably made on the basis of a high molecular weight PACA (homo)polymer, is in the micrometer range, preferably less than 700 μm, more preferably less than 500 μm, more The average fiber diameter of the mesh having a thickness more preferably less than 400 μm, even more preferably less than 300 μm may range between 500 nm and 1.5 μm, the mesh also having a thickness in the range of about 0.5 MPa to 1.5 MPa. Young's modulus, tensile strength ranging from about 0.05 MPa to 0.15 MPa. Also, such a mesh may have a strain to failure of about 15%-25%. In an even more preferred embodiment, a mesh as disclosed herein, preferably a mesh made on the basis of a high molecular weight PACA (homo)polymer, may have a thickness in the range ≤ 400 μm, the average of such meshes being The fiber diameter is in the range between 500 nm and 1.5 μm, preferably between 500 nm and 1.5 μm. More preferably, such a mesh may also have a Young's modulus in the range of about 0.5 MPa to 1.5 MPa, preferably in the range of about 0.7 MPa to 1.2 MPa. In an even more preferred embodiment, such a mesh may also have a tensile strength in the range of about 0.05 MPa to 0.15 MPa, preferably in the range of about 0.06 MPa to 0.1 MPa. Also, such a mesh may have a strain to failure of about 15%-25%.

Table 2 below shows the physical and mechanical properties of the electro-spun mesh encompassed by the present disclosure.

The tensile properties of a mesh composed of PACA having a number average molecular weight of the order of 10 ^{6 Da may be as follows: Young's modulus = 0.3-3 MPa and tensile strength = 0.02-0.2 MPa.} This mesh is stable for at least 23 days in culture medium containing human gingival fibroblasts. 10 is a SEM picture of a PBCA mesh before and after 23 days of incubation with FHGFib.

In various embodiments of the present disclosure, a mesh as disclosed herein, preferably a nano/microfiber mesh prepared on the basis of a high molecular weight PACA (homo)polymer as described in the preceding paragraph, is from 0.1 μm ² to It may have a pore area in the range of 500 μm ^{2 .} Preferably, such a mesh comprises a high molecular weight PACA polymer, preferably a high molecular weight PACA homopolymer (such as PBCA) and a non-PACA polymer, preferably a non-PACA homo- or copolymer (such as chitosan or PLGA, respectively) It can be based on electrospinning of a blend solution of In various other embodiments of the present disclosure, a mesh as disclosed herein, preferably a nano/microfiber mesh made on the basis of a high molecular weight PACA (homo)polymer as described in the preceding paragraph, is 0.1 μm ² to 370 μm ² , in which case the mesh does not contain a non-PACA polymer or therapeutic agent and is instead of a solution containing only a high molecular weight PACA polymer, preferably a high molecular weight PACA homopolymer (such as PBCA). It can be based on electrospinning.

In various embodiments, the ready-to-use poly(alkyl cyanoacrylate)-based nano/microfibers (and/or PACA-based nano/microfiber meshes) disclosed herein have a polydispersity index ranging from 1 to 1.7. obtained from PACA polymers. Surprisingly, the ready-to-use poly(alkyl cyanoacrylate)-based nano/microfibers (and/or PACA-based nano/microfiber meshes) were obtained from PACA polymers having a polydispersity index outside the range of 1 to 1.7. It has been found by the present inventors that it has excellent mechanical properties when compared to Preferably, the excellent mechanical properties of the nano/microfiber mesh are such that it is between 600 and 700 μm, between 500 and 600 μm, between 400 and 500 μm, between 300 and 400 μm, between 200 and 300 μm and/or 100 μm. It is characterized in that it can have a thickness in the range of micrometers between to 200 μm. In a specific embodiment, the thickness is approximately 431 μm. In another specific embodiment, the thickness is approximately 451 μm. In another specific embodiment, the thickness is approximately 411 μm. In another specific embodiment, the thickness ranges from 411 to 451 μm. Preferably, the excellent mechanical properties of the nano/microfiber mesh also indicate that it is in the range of 100 nm to 5 μm, between 5 and 4 μm, between 4 and 3 μm, between 2 and 1 μm, between 1000 and 900 nm, 900 a fiber diameter between 800 nm, 800 and 700 nm, 700 and 600 nm, 600 and 500 nm, 500 and 400 nm, 400 and 300 nm, 300 and 200 nm, 200 and 100 nm. It is characterized by being able to have In a specific embodiment, the thickness is approximately 1 μm. In another specific embodiment, the thickness is approximately 1.3 μm. In another specific embodiment, the thickness is approximately 0.7 μm. In another specific embodiment, the thickness ranges from 0.7 to 1.3 μm. Preferably, the excellent mechanical properties of the nano/microfiber mesh also indicate that it is in the range of 0.2 to 4 MPa, between 0.2 and 0.5 MPa, between 0.5 and 0.8 MPa, between 0.8 and 1 MPa, between 1 and 1.2 MPa, between 1.2 and 1.2 MPa. It can have a Young's modulus between 1.4 MPa, between 1.4 and 1.8 MPa, between 1.8 and 2.4 MPa, between 2.4 and 3 MPa, between 3 and 3.4 MPa, between 3.4 and 3.8 MPa, between 3.8 and 4 MPa. . In a specific embodiment, the Young's modulus is approximately 1.4 MPa. In another specific embodiment, the Young's modulus is approximately 1.45 MPa. In another specific embodiment, the Young's modulus is approximately 1.35 MPa. In another specific embodiment, the Young's modulus ranges from 1.35 to 1.45 MPa. Preferably, the excellent mechanical properties of the nano/microfiber mesh also indicate that it is in the range of 0.02 to 0.3 MPa, between 0.02 and 0.05 MPa, between 0.05 and 0.08 MPa, between 0.08 and 0.1 MPa, between 0.1 and 0.12 MPa, between 0.12 and 0.12 MPa. It may have a tensile strength of between 0.14 MPa, between 0.14 and 0.18 MPa, between 0.18 and 0.24 MPa, between 0.24 and 0.3 MPa. In a specific embodiment, the tensile strength is approximately 0.09 MPa. In another specific embodiment, the tensile strength is approximately 0.07 MPa. In another specific embodiment, the tensile strength is approximately 0.05 MPa. In another specific embodiment, the thickness ranges from 0.05 to 0.09 MPa. In a separate embodiment, a ready-to-use poly(alkyl cyanoacrylate) based nano/microfiber obtained from a PACA polymer having a polydispersity index in the range of 1 to 1.7 with the excellent mechanical properties of the above embodiment (and / or PACA-based nano/microfiber mesh) is preferably obtained from high molecular weight PACA. In a preferred embodiment, the high molecular weight is a number-average molecular weight.

In a separate embodiment, the ready-to-use poly(alkyl cyanoacrylate)-based nano/microfibers (and/or PACA-based nano/microfiber meshes) with excellent mechanical properties are 2.0 or less, 2.4 or less, 2.5 or less; It may be obtained from a PACA polymer having a polydispersity index in the range of 2.7 or less, or in the range of about 1.8 to 2.4, about 1.8 to 2.5, about 1.8 to 2.7, about 2.4 to 2.7. In another embodiment, the ready-to-use poly(alkyl cyanoacrylate) based nano/microfibers (and/or PACA based nano/microfiber mesh) with excellent mechanical properties are preferably 2.0 or less, 2.4 or less; It is obtained from a high molecular weight PACA polymer having a polydispersity index in the range of 2.5 or less, 2.7 or less, or in the range of about 1.8 to 2.4, about 1.8 to 2.5, about 1.8 to 2.7, about 2.4 to 2.7. In various embodiments, the molecular weight is a number-average molecular weight. In various embodiments, the polydispersity index refers to the ratio of the weight-average molecular weight (M _{w ) to the number-average molecular weight (M n} ) of the PACA polymer of a PACA homopolymer and/or a PACA copolymer.

As exemplified herein, a multilayer electro-spun biodegradable hydrophobic mesh composed of PACA/PLGA random nano/microfibers with good adhesion between nanofiber layers and a porous morphology is obtained. Such a mesh may aid in cell adhesion and proliferation, and aid in tissue repair by virtue of its hemostatic and bacteriostatic properties. A mesh based on PACA/PLGA can be composed of a mixture of ribbon-like nanofibers and full cylinder-like nanofibers (shown in FIG. 6 ). The thickness of such a mesh may generally be less than 500 μm. The average fiber diameter may range from 800 nm to 2 μm. The porosity of the mesh may be between 30% and 50%. Its mechanical properties may be as follows: Young's modulus = 0.2-4.0 MPa, tensile strength = 0.02-0.2 MPa.

In addition, a multilayer electro-spun biodegradable mesh composed of PACA and chitosan random nano/microfibers with good adhesion between nanofiber layers and a porous morphology is obtained. These meshes are used for cell adhesion and proliferation and may aid in tissue repair by virtue of their hemostatic and bacteriostatic properties. A PACA/PACA-co-CS-50% mesh composed of a mixture of ribbon-type nanofibers and filled cylindrical nanofibers may have an average fiber diameter of about 1 μm and a porosity of about 70%. The thickness of this mesh may be less than 500 pm. Its mechanical properties may be as follows: Young's modulus = 0.8 ± 0.3 MPa, tensile strength = 0.06 ± 0.02 MPa.

Electrospinning of a PACA-based solution—any polymer solution containing PACA—can be used to prepare meshes for potential biomedical applications. Using bulk anionic polymerization of the corresponding monomers with an anionic initiator having a weight-average molecular weight in the range of 10 ⁶ -10 ⁴ _{Da, PACAs with different number-average molecular weights (M n} ) were synthesized (10 ^{6 ).} Da, 10 ⁵ Da, 10 ⁴ Da, 10 ³ Da).

Filled with, immobilized with, or encapsulated with particles of a therapeutic agent/other chemical or a biodegradable polymer filled with such therapeutic/other chemical having good adhesion and porous morphology between nanofiber layers. A multilayer electro-spun biodegradable mesh composed of PACA random nano/microfibers can be obtained. Such meshes can be used for cell adhesion and proliferation, aid in tissue repair by their hemostatic and bacteriostatic properties, and can be used to deliver therapeutics/other chemicals. The mesh consisted of homogeneous nano/microfibers in which PACA and other components were homogeneously mixed in a common solvent and electro-spun (see FIG. 6 ). The thickness of this mesh was less than 500 μm, with an average fiber diameter ranging from 200 nm to 2 μm and a porosity between 20 and 70%. The mechanical properties of one of the formulations (PBCA/5FU_15%) were: Young's modulus = 0.4 ± 0.2 MPa, tensile strength = 0.08 ± 0.04 MPa.

The novel PACA-based nano/microfiber mesh of the present invention may be provided in a rectangular shape such as, for example, 10×10 cm. The rectangle may then be cut into a shape deemed more appropriate for the intended application. Advantageously, since the PACA-based nano/microfiber mesh of the present invention is itself adherent to the skin, it is not necessary to apply the mesh to an adhesive or support. Nevertheless, if deemed advantageous, an adhesive, such as a cyanoacrylate adhesive, may be applied to adhere the edges of the mesh to the site of application to further improve adhesion.

The porosity of the novel PACA-based nano/microfiber mesh of the present invention is preferably between 40 and 70%, more preferably between 50 and 70%. The mesh is effective, for example, as a protective barrier that inhibits the entry of exogenous microorganisms into the wound. In contrast to films formed from PACA, which is generally a non-porous material, the mesh is characterized by being highly porous with pores that are well interconnected, which makes it possible, for example, to exudate fluid from a wound. , allowing gas permeation, and cell proliferation and migration. The fibrous mesh of the present invention is advantageous over films of the same PACA material as it has a greater surface area for cell adhesion and cell proliferation. As described elsewhere herein, the porous mesh provides a high surface area-to-volume ratio, can help control fluid drainage, and can be used as a delivery platform for therapeutic agents, other chemical or biological materials. . Porosity can generally be measured by porosimetry, by measuring the apparent material density, by BET analysis, or by microscopic imaging. Porosity can be measured using BET analysis using nitrogen as the adsorbate. Porosity values may also be obtained by analysis of SEM pictures using ImageJ software.

Within 30 days in PBS medium, consistent drug release patterns of up to 51-55% were observed. 9 shows the release of 5FU from PACA/5FU 15% mesh incubated in PBS pH=7.4 at 37°C. At day 96, drug release was 80-88% of the total drug content in all samples (see Figure 9).

Multilayer electro-spun biodegradable meshes can be provided that are composed of PACA random nano/microfibers filled with any type of animal cells, including human cells, which may belong to the subject itself being treated. Such meshes can be effectively used in tissue engineering.

As described elsewhere herein, the PACA polymer concentration, particularly the PBCA concentration, may preferably be between 0.5 and 50 % w/w in acetone or an acetone/ethanol mixture. In various embodiments, the PACA is ^{a PACA homopolymer, particularly a PBCA homopolymer, having a low molecular weight, such as 10 3} Da, the polymer concentration (PACA, particularly PBCA) being between 20% w/w and 50% w/w in solution. is the range of Preferably, the molecular weight represents a number-average molecular weight. In various preferred embodiments, the solvent is acetone or an acetone/ethanol mixture. Preferably, the polymer concentration (PACA, in particular PBCA) of the low molecular weight polymer ranges between 30 % w/w and 50 % w/w, wherein the solvent is preferably acetone or an acetone/ethanol mixture. More preferably, the polymer concentration (PACA, especially PBCA) of the low molecular weight polymer ranges between 35 % w/w and 50 % w/w, wherein the solvent is preferably acetone or an acetone/ethanol mixture. Even more preferably, the polymer concentration (PACA, especially PBCA) of the low molecular weight polymer ranges between 40 % w/w and 50 % w/w, wherein the solvent is preferably acetone or an acetone/ethanol mixture. In a particularly preferred embodiment, the low molecular weight polymer concentration (PACA, in particular PBCA) is about 50 % w/w, wherein the solvent is preferably acetone or an acetone/ethanol mixture. Such embodiments may provide a more viscous PACA solution.

As disclosed herein, depending on the solvent used, fibers having different cross-sectional areas can be produced. Specifically, the method of the present invention makes it possible to process PACA materials into different nanofiber morphologies, including ribbon-like (flat) or filled cylindrical (circular), without altering the PACA chemical structure. For example, when acetone/ethanol is used, round fibers can be produced, and when acetone is used alone, flat fibers can be produced.

Advantageously, since PACA is biodegradable, nano/microfiber meshes are formed from PACA materials that are degradable, for example by the action of various enzymes such as esterases under the normal functioning of the human body and living organisms, in which case the by-products are The corresponding alkyl alcohols and polycyanoacrylic acids which are water-soluble substances and which are removed in vivo by renal filtration. As such, the mesh can be used in a variety of applications where the mesh completely disintegrates within the body over time without the need for the mesh to be removed after use, for example by further surgery. Especially in the case of dental applications, the omission of the second operation for removal of the mesh is advantageous to ensure a better healing process of the tissue being treated. In addition, PACAs are generally bacteriostatic and can stop the growth of Gram-positive and some Gram-negative bacteria. PACA is also hemostatic. Thus, the implanted mesh can be used to cause a mechanical blockage to slow blood flow, or to provide a surface agent that activates the coagulation cascade. When used as a tissue adhesive, the porous mesh can advantageously allow blood to penetrate with subsequent coagulation into the pores of the mesh.

In some aspects, the present disclosure also provides a method of treating or preventing a disease or disorder, for example, by topical administration of novel ready-to-use poly(alkyl cyanoacrylate)-based nano/microfibers as described herein. , or a method of maintaining a healthy or excellent condition. For example, in some embodiments, the present disclosure provides a method of treating an external wound, the method comprising: a novel ready-to-use poly(alkyl cyanoacrylate) based as disclosed herein. Topically administering nano/microfibers to an external wound on a subject. As another example, the present disclosure provides a method of treating an external burn, the method comprising: a novel ready-to-use poly(alkyl cyanoacrylate) based nano/microfiber as disclosed herein Topically administering to an external burn of a subject. As another example, the present disclosure provides a method for the treatment of aches and pains, the method comprising: a novel ready-to-use poly(alkyl cyanoacrylate) based nano/micro as disclosed herein Topically administering a fiber to a skin region of a subject, wherein the skin region is preferably proximate (eg directly over) the pain or pain. As another example, the present disclosure provides a method of promoting gingival health (eg, by inhibiting the growth of oral bacteria), the method comprising: a novel ready-to-use as disclosed herein Oral administration of possible poly(alkyl cyanoacrylate)-based nano/microfibers to a subject. As another example, the present disclosure provides a method of treating peri-implantitis, the method comprising: a novel ready-to-use poly(alkyl cyanoacrylate) based nano/micro as disclosed herein Orally or dentally administering the fiber to the subject.

In some embodiments of the present invention, the ready-to-use PACA-based nano/microfibers contemplated herein may be suitable for uses such as cosmetic and/or personal care applications for: (i) assisting in moisture control; coating and protection as film-forming components; (ii) providing antimicrobial activity as a natural preservative; and/or (iii) acting as a carrier for other active ingredients for slow release due to ionic binding to the compound and degradation of chitosan by natural skin enzymes.

In some embodiments of the present invention, the ready-to-use PACA-based nano/microfibers contemplated herein can be applied in a variety of ways, for example: (i) covering a wound or damaged/inflamed skin or area. , sealing and/or protecting; (ii) creating a customizable, smooth film that can be degraded by skin enzyme(s) and that is readily formed to conform to the skin or area upon treatment; (iii) providing suitable conditions for moisture control; (iv) inhibiting skin and wound infections; (v) reducing itching and inflammation; (vi) contributing to rapid healing; and/or (vii) reducing hypertrophic scarring; (viii) alleviating aches and pains associated with muscle and joint ulcers; and/or (ix) treating disorders within the oral mucosa and dental supporting tissue. Additional uses contemplated herein include targeted cleaning of dirty wounds, breast care of milking animals, treatment of human and animal wounds or burns, soothing skin inflammation, alleviating muscle aches and pains, and/or insect bites. or to reduce itching due to plant toxins, sunburn, bedsores, ulcers, for example, to promote oral health by inhibiting the growth of oral bacteria, and to treat other disorders of the oral mucosa and tooth-supporting tissues including doing. The ready-to-use PACA-based nano/microfibers contemplated herein have a number of advantages, for example, a mesh of nano/microfibers results in a thicker film that lasts longer than a spray solution; Such materials are less prone to rapid biodegradation, adapt more readily to wounds than bandages, cover only the required areas, and can be custom transparent flexible "bandages" with a variety of properties.

In some aspects, the present disclosure provides for the promotion of oral health, comprising orally applying (or administering) any of the ready-to-use PACA-based nano/microfibers disclosed herein to the soft or hard tissue of a subject's mouth. (e.g., promotion of gingival health). In a similar embodiment, the present disclosure provides any of the ready-to-use PACA-based nano/microfibers disclosed herein for use in promoting oral health (eg, promoting gingival health). In another similar embodiment, the present disclosure provides for the manufacture of a medical device or medicament for promoting oral health (such as promoting gingival health) of any of the ready-to-use PACA-based nano/microfibers disclosed herein. provide use. The present disclosure relates to the treatment of disorders of oral mucosa and/or dental support tissue, such as treating peri-implantitis, comprising orally applying or administering to a subject any of the ready-to-use PACA-based nano/microfibers disclosed herein. provide a way

The present invention specifically relates to the novel PACA-based nano/microfibers and/or PACA-based nano/microfiber meshes disclosed herein for wound dressing/healing, tissue regeneration/engineering, organ protection, implant coating or controlled drug delivery. Use for use in dental and/or oral applications is encompassed. Said use of the novel PACA-based nano/microfibers and/or PACA-based nano/microfiber meshes of the invention in dental and/or oral applications is a preferred embodiment.

The present invention specifically relates to the use of the novel composite materials disclosed herein for use in dental and/or oral applications, for wound dressing/healing, tissue regeneration/engineering, organ protection, implant coating or controlled drug delivery. covers Said use of the novel composite material of the invention in dental and/or oral applications is a preferred embodiment. In various preferred embodiments, the composite material comprises (i) a PACA-based nano/microfiber or PACA-based nano/microfiber mesh disclosed herein and (ii) chitosan.

A patient or subject to be treated by any of the novel ready-to-use poly(alkyl cyanoacrylate) based nano/microfibers or methods of the present disclosure may refer to either a human or a non-human animal. In one embodiment, the present disclosure provides a method of treating a disease in a human patient in need thereof. In one embodiment, the present disclosure provides a method of treating an inflammatory disease in a human patient in need thereof. In another embodiment, the present disclosure requires those including, but not limited to, dogs, horses, cats, rabbits, gerbils, hamsters, rodents, birds, aquatic mammals, cattle, pigs, camels and other zoological animals. It provides a method of treating a disease in a veterinary patient.

The term “treating” refers to: a disease, disorder or condition occurring in a cell, tissue, system, animal or human that is predisposed to the disease, disorder and/or condition but may not yet have been diagnosed as having it. to prevent; stabilizing the disease, disorder or condition, ie, arresting its development; and/or alleviating one or more symptoms of the disease, disorder or condition, ie, causing regression of the disease, disorder and/or condition. As used herein, a therapeutic agent that “prevents” a disorder or condition reduces the incidence of the disorder or condition in a statistical sample in a sample to be treated compared to an untreated control sample, or one or more of the disorder or condition compared to an untreated control sample. Refers to a compound that delays the onset of or reduces the severity of symptoms. Accordingly, in some aspects and embodiments of the present disclosure, treatment of a disease or condition comprising topical administration of novel ready-to-use poly(alkyl cyanoacrylate) based nano/microfibers as disclosed herein or a method of prevention is provided.

The word "comprises/comprising" and the word "having/comprising" when used herein in reference to the present invention are used to specify the presence of the stated feature, integer, step or element, but not other It does not exclude the presence or addition of one or more features, integers, steps, elements or groups. In various embodiments, the term “comprising” may encompass the term “consisting of”.

Also, the term “about” is known to one of ordinary skill in the art in the context in which it is used herein. Specifically, "about" is meant to refer to and refer to variations that are ±20%, ±10%, preferably ±5%, more preferably ±1%, even more preferably ±0.1% of the indicated value. It is also understood that certain features of the invention, which, for clarity, are described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which, for brevity, are described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination.

From now on, the present disclosure will be described in detail with reference to the following examples. However, the examples are for illustrative purposes only and are not intended to limit the scope of the present disclosure.

[ Example ]

In the examples below, the resulting polymer solutions and emulsions were placed in horizontally oriented 10 mL plastic syringes coupled with needles with different inner diameters from 25G to 15G. The following different collectors were used: copper mesh, copper ring, aluminum foil, paper and stainless steel squares. The collector was grounded and placed at a distance of 5-30 cm from the needle. The voltage applied to the needle was in the range of 5-30 kV. Flow rates ranged from 0.025 to 2.000 mL/min. Scanning electron microscopy (SEM) was used to confirm the morphology of the resulting mesh. The pore properties and average fiber diameter of the nonwoven mesh were determined from SEM photographs using ImageJ software version 1.51n (http://imagej.nih.gov/ij). First, a mesh based on PACA electro-spun nano/micro-fibers was sputter coated with gold using a Balzers Sputter Coater SCD 040 (Manchester, NH) for 2 minutes and then coated with gold by Maxim (Cambridge, UK). ) Sputtering coated meshes were observed using a scanning electron microscope at a working distance between 18-30 mm and an accelerating voltage of 15-20 kV.

10³-10⁴ Da)을 갖는 PACA의 경우이다. 도 13은 제조 시점 ("0년") A1) 및 B1); 및 정상 조건에서 1년 저장 후의 A2) 및 B2)로 측정된 PLGA 및 PACA/PLGA를 기재로 하는 메시의 SEM 사진이다.By preparing solvent mixtures of different combinations, generally ribbon-like electro-spun fibers ( FIG. 12 , A1 ) and filled cylindrical electro-spun fibers ( FIG. 6 , B1 ) were obtained. 12 shows A1) flat fibers and B1) round fibers at the time of manufacture (“Year 0”); and A2) and B2) mesh SEM pictures based on PACA after 1 year of storage under normal conditions. The fiber diameter was mainly controlled by the concentration of the PACA-based solution (0.1-70% (w/w)), the smaller for PACA with a number average molecular weight between ^{10 5} and 10 ^{6 Da, and the larger one} lower number average molecular weight (

10 ³ -10 ⁴ Da). 13 shows the time of manufacture (“year 0”) A1) and B1); and SEM pictures of meshes based on PLGA and PACA/PLGA measured in A2) and B2) after 1 year of storage under normal conditions.

Example One

Electrospinning of PHCA-poly(hexyl cyanoacrylate)

A mixture of acetone/ethanol 2/1 (v/v) was prepared. A 0.1% (w/w) solution was prepared by dissolving PHCA having a number average molecular weight of about ^{10 6 Da in a solvent mixture using magnetic stirring.} Place the solution inside a plastic 10 mL syringe coupled to a stainless steel needle tip 25G. A collector ( ^{a square copper mesh of 100 cm 2} ) is placed at a distance of 10 cm from the needle tip. The flow rate is fixed at 0.125 mL/min, and 10 kV is applied for a period of 30 min. Cylinder-shaped filled nanofibers with an average fiber diameter of 696±113 nm, a porosity of 54% and a thickness of less than 500 μm are obtained. The mechanical properties are summarized in Table 2, with a Young's modulus of 1.2 MPa and a tensile strength of 0.08 MPa.

Example 2

Electrospinning of POCA and PLGA (POCA = poly(octyl cyanoacrylate))

A blend of POCA and PLGA powder with a PLGA maximum concentration of 50% or less was dissolved in ethyl acetate. ^{POCA and PLGA in a 1/1 ratio with a number average molecular weight of the order of 10 6} Da were prepared in the following two alternative ways: 1) one having a POCA concentration of 0.5% and one having a PLGA concentration of 12% preparing the two polymer solutions separately and then mixing them; 2) preparing a PLGA solution; After PLGA is completely dissolved, POCA is added, followed by stirring until the added POCA is completely dissolved. The resulting solution was placed inside a plastic 10 mL syringe coupled to a stainless steel needle tip 17G. A collector (10 cm ² of square-shaped aluminum foil) was placed at a distance of 7 cm from the needle tip. The flow rate was fixed at 0.5 mL/min. 20 kV was applied for a period of 30 minutes. A mixture of cylindrical and ribbon-shaped nanofibers was obtained, and the fiber diameter was about 900 nm. These meshes are thicker than those based on POCA alone, but have a fiber diameter of less than 500 nm. The porosity of the mesh was measured to be about 39%. The mesh had a Young's modulus of about 1 MPa and a tensile strength of about 0.06 MPa.

Example 3

Electrospinning of POCA and PIBCA-co-chitosan

Meshes based on PACA and chitosan were prepared in two alternative ways: 1) emulsion by dissolving chitosan having a weight average molecular weight of the order of ^{10 6 Da in 1% in acetic acid at 0.1 mol/L} to obtain, and separately preparing a solution of isobutyl cyanoacrylate (IBCA) at 2% in acetone. The IBCA solution was added to the chitosan solution while maintaining the IBCA/chitosan combination at 60/40 (w/w). When polymerization was complete, acetone was added to the emulsion to dilute it. The emulsion was stirred overnight. One drop of a dense portion of the emulsion was diluted in butanol, and this fresh emulsion was used for electrospinning (voltage = 15 kV; needle tip-collector distance = 5 cm; needle tip = 18 G; flow rate = 0.250 mL/min). . 2) POCA was mixed with copolymer PIBCA-co-chitosan (prepared in the above-mentioned emulsion system) while maintaining the POCA/PIBCA-co-chitosan combination at 70/30 w/w, where total polymer in acetone % was 1%.

The mesh produced by alternative 1) resulted in a sponge-like structure with some particles having a diameter between 0.9 and 8 mm (Fig. 6,E). The mesh prepared by alternative 2) had a fiber diameter of about 1 μm and a thickness of about 400 μm, a porosity of 69%, a Young's modulus of about 1 MPa, and a tensile strength of about 0.07 MPa.

Example 4

Electrospinning of PPCA and drug (PPCA = poly(n-pentyl-2-cyanoacrylate)

Prepare a solution of 5FU in formic acid. PPCA having a molecular weight of 10 ⁵ Da was added to obtain a 2% (w/w) solution, and 5FU was 15% based on the weight of PPCA. The polymer solution was placed inside a plastic 10 mL syringe coupled to a stainless steel needle tip 21G. The collector ( ^{square-shaped stainless steel of 1 cm 2 on} aluminum foil) was placed vertically at a distance of 20 cm from the needle tip. The flow rate was fixed at 0.05 mL/min and 22 kV was applied. A mixture of cylindrical nano/microfibers and ribbon nano/microfibers was obtained, with an average fiber diameter of 798 ± 310 nm and a porosity of 60%. The thickness of the mesh was about 390 pm, the Young's modulus was about 0.7 MPa, and the tensile strength was about 0.1 MPa.

Example 5

Electrospinning of PBLCA-poly(butyl-lactoyl-2-cyanoacrylate)

PBLCA having a number average molecular weight of about 10 ³ Da was dissolved in a dimethylformamide/acetone = 1/2 (v/v) solvent mixture using magnetic stirring to prepare a 50% (w/w) solution. The resulting solution was placed inside a plastic 10 mL syringe coupled to a stainless steel needle tip 15G. A collector (10 cm ² square-shaped aluminum foil) was placed at a distance of 5 cm from the needle tip. The flow rate was fixed at 2 mL/min and 13 kV was applied for a time period of 30 min. Cylindrical and very well connected fibers having an average fiber diameter of 1320±721 nm, a porosity of 41% and ^{an average pore area of 11 μm 2} were obtained ( FIG. 6A ).

Example 6

Electrospinning of PHepCA-D&C Violet #2 Solution (PHepCA = poly(n-heptyl-2-cyanoacrylate))

In acetone/ethanol = 2/1 and acetone, respectively, PHepCA and D&C Violet #2 solutions were prepared separately. The D&C Violet #2 solution was 8 pg/mL and the PHepCA solution (with ^{a weight average molecular weight in the range of 10 4} Da) was 20% (w/w). One drop of the staining solution was added to the PHepCA solution and stirred until the final solution was homogeneous. The solution was placed inside a plastic 10 mL syringe coupled to a stainless steel needle tip 20G. A collector (a copper ring with a diameter of 1 cm) was placed at a distance of 15 cm from the needle tip. The flow rate was fixed at 1 mL/min and 11 kV was applied for a period of 30 min. Nanofibers were obtained in different layers. Using an LSM 700 laser scanning microscope in fluorescence mode (Carl Zeiss Microscopy GmbH, Jenna 07745, Germany), the obtained multilayer assembly was confirmed. 13 shows confocal laser scanning microscopy of PACA mesh stained with D&C Violet #2.

Example 7

Electro-spun PBCA scaffolds for cell culture (support for culturing/growing cells, especially human cells)

Using a stainless steel rectangular copper mesh attached as a collector, from electrospinning of a PBCA solution having a weight average molecular weight ^{of 10 6} Da, 1% in acetone/ethanol = 3/2 (v/v), 0.025 mL/min. A mesh was obtained with a feed rate, 5 kV, and a collector distance of 5 cm. This mesh was composed of PBCA nano/microfibers in the shape of a filled cylinder with an average diameter of 1.0 ± 0.4 μm.

Meshes were used in different experiments. FIG. 5 shows fluorescence photographs of human gingival fibroblasts growing on PBCA meshes during 14 days of incubation at 37° C. and 5% (v/v) CO _{2 .} Assays were performed after 3, 7, 14 and 23 days of HGFib incubation on PBCA mesh. As can be seen, the cells spread after 7 days, and after 14 days more than 70% of the mesh area was occupied by the living and proliferating cells.

Cell adhesion as a result of seeding was measured as follows: To assay for adherent cells, the medium was removed after days of incubation (3, 7 and 14) and live/dead using fluorescein and ethidium bromide, respectively. By performing staining, a fluorescence photograph using an Olympus fluorescence microscope VANOX-T was obtained. The fluorescence photograph (see Fig. 5) shows that the mesh composed of PBCA is capable of culturing human gingival fibroblasts.

Example 8

Measuring the mechanical properties of the mesh

A uniaxial tensile test (Zwick/Z010, Germany, equipped with a 1000 N load cell) was used to determine the mechanical properties of the samples. Specimens were prepared by cutting the samples into rectangular strips 10 mm wide by 13 mm long (gauge length of 10 mm) with a thickness range of 300-400 μm. The crosshead speed was set at 10 mm/min and the analysis was performed at ambient conditions. At least three samples were analyzed for each nanofiber mesh type. Tensile strength and Young's modulus were derived from stress-strain curves generated from force versus elongation data measured by the testing machine (Tables 2 and 3).

Example 9

Measuring the contact angle of the mesh

The surface wettability of the electro-spun mesh was detected by the distilled water contact angle method at room temperature. Water was dripped onto the mesh (on at least 3 different places) and one picture was taken immediately using optical microscopy (KEYENCE VHX-5000, Germany), followed by subsequent use of the Keyence software. The water contact angle was determined by imaging the floating droplet (Tables 2 and 3).

Example 10

Use of mesh as a membrane

As outlined above, a nanofiber mesh of PBCA was obtained. After closing the bone defect, a membrane was formed from the PBCA mesh by cutting it into the required shape and placing it as a membrane between the hard and soft tissues.

Example 11

As outlined above, a nanofiber mesh of PBCA was obtained. The mesh was formed into a required shape and used as a sleeve for dental implants.

Example 12

The surface of the biodegradable Mg alloy biomaterial was coated using the PBCA nanofiber mesh obtained as outlined above. This allowed the decomposition rate of the Mg alloy to be controlled, thereby reducing hydrogen production and thus affecting the decomposition. In addition, an almost stable decomposition rate could be achieved.

Example 13

Tests for cell adhesion and proliferation (potential tissue regeneration/engineering scaffold utility) were performed as follows: First, human gingival fibroblasts were ^{plated as monolayers in two 25 cm 2} tissue culture flasks, and cells were Incubate until passage. After passage 5, cells were removed by trypsinization, counted and seeded onto meshes at a density of ^{37,500 cells/cm 2 .} To inactivate possible microbial contaminants that may have arisen during the electrospinning process, the mesh was first exposed to sterile ultraviolet radiation for 30 minutes (sterilization procedure, FIG. 9 ). Cells in contact with the mesh were maintained at 37° C. with _{5% CO 2 in an incubator.} The medium was changed every 3 days.

Example 14

Preparation of PACA by Anionic Mass Polymerization - Synthesis of PBCA with a molecular weight greater than ^{10 3 Da}

을 갖는 PBCA를 제조하였다. 반응은 저온 수조상 3-목 원형-저 플라스크에서 수행하였다. BCA를 먼저 플라스크에 첨가한 후, 이어서 개시제 (아세톤 중에 희석된 것)를 첨가하였다. 플라스크를 약하게 교반하였다. 중합 종료시 (고체 형성, 저온 반응 시스템, 가시적인 변화 없음), 고체를 용해시키고, 증류수에서 침전시켜, 여과하고, 먼저 물로 세척한 후 나중에 메탄올로 세척하고, 진공에서 건조하였다. 하기 표 1은 해당

값 및 BCA와 DMPT 사이의 몰 조합을 갖는 PBCA의 확인을 보여준다.By bulk anionic polymerization of BCA using DMPT as an initiator, about 10 ⁴ , 10 ⁵ and 10 ⁶ Da

PBCA with The reaction was carried out in a 3-neck round-bottom flask on a cold water bath. BCA was first added to the flask, followed by the initiator (diluted in acetone). The flask was gently stirred. At the end of the polymerization (solid formation, low temperature reaction system, no visible change), the solid was dissolved, precipitated in distilled water, filtered, washed first with water, then with methanol and dried in vacuo. Table 1 below is

values and identification of PBCAs with a molar combination between BCA and DMPT.

Example 15

Mesh based on PBCA alone:

With respect to PBCA molecular weight, electro-spun PBCAs with different molecular ^{weights from high molecular weight (10 6} Da) to low molecular weight (10 ³ Da) with a polymer concentration of 0.5 to 50 % w/w in acetone or acetone/ethanol mixture. Could. The thickness of the mesh was about 260 μm. The fiber diameter of the mesh increases with the concentration of polymer. For example, using 0.5% polymer solution (PBCA high molecular weight), PBCA nanofiber-based mesh (600 nm) was obtained, whereas using 1.1% w/w polymer solution (PBCA high molecular weight) microfiber mesh was used. (about 1 μm) was obtained. Surprisingly, it has been tested that continuous smooth fibers can be obtained using this very low viscosity PBCA solution (0.3-3 cP), which is not possible with other polymers such as PCL. For PBCA with low molecular weight, a viscous solution (up to 50 % w/w) was used for electro-spinning. When acetone/ethanol is used, fibers with different cross-sectional areas can be produced, which are circular, and flat fibers when acetone is used alone. The pore area of the mesh ranges ^{from 0.1-370 μm 2 .} When the fibers were flat or circular, the mechanical properties of the mesh composed of PBCA with high molecular weight were not so much differentiated, depending on the aging of the mesh (at least 1 year) as well as depending on the range of fiber diameters tested (600 nm and 1.5 μm) were not. The mesh had a Young's modulus of about 1.2 MPa, a tensile strength of about 0.08 MPa, a strain to break of about 15%, and the values were closer to elastic materials than rigid materials.

Example 16

Mesh based on PBCA-PLGA:

The mesh of this example was fabricated by electrospinning a blend solution of PBCA high molecular weight and PLGA (75:25 high molecular weight powder having a PLGA maximum concentration of 50% or less in acetone with a maximum concentration of PBCA 1% w/w). prepared. The blend solution was prepared using the following two methods: 1) separately preparing a polymer solution in two acetone, one PBCA 1% and the other PLGA 75:25 12%, and then mix them; 2) Prepare PLGA acetone solution, dissolve PLGA, add PBCA, and stir until PBCA is completely dissolved. These meshes were thicker (about 390 μm) compared to those based on PBCA alone. The fiber diameter was about 900 nm. The pore area of the mesh was in the range of ^{0.1-160 μm 2 .} The mesh had a Young's modulus of about 1 MPa, a tensile strength of about 0.06 MPa, and a breaking strain of about 20%.

Example 17

PBCA-chitosan based mesh

Based on PBCA and chitosan by mixing PBCA with high molecular weight with PBCA-chitosan copolymer (powder) prepared in the emulsion system previously described while maintaining the PBCA/PBCA-chitosan combination at 50/50 w/w. A mesh was prepared, wherein the % total polymer in acetone was 2%. The fabricated mesh had a fiber diameter of about 1 μm, a thickness of about 400 μm, ^{a minimum pore area of 0.1 μm 2} , a maximum pore area of 482 μm ² , a Young's modulus of about 1 MPa, a tensile strength of about 0.07 MPa and a 14% had a breaking strain.

Example 18

Mesh based on PBCA-5FU

Different concentrations of drug 5FU were incorporated in the polymer solution for electrospinning: 7, 10, 15, 20, 25 and 30% by mass of the high molecular weight PBCA used in 1 and 2% acetone. It was shown that the drug was homogeneously incorporated inside the fibers (from 700 nm to 1.2 μm) in such a way that the fiber diameter also increased with increasing drug concentration, and this was tested in a kinetic release assay at 37 °C in PBS medium. A consistent drug release pattern was observed until day 98 of incubation. The mesh composed of PBCA with high molecular weight and 15% of 5FU had a thickness of about 390 μm, and as a result of tensile testing, Young's modulus of about 0.7 MPa, tensile strength of about 0.1 MPa, and breaking strain of 25% were obtained.

Example 19

Mesh based on PBCA-PLA-Mg

PBCA/PLA-Mg combinations in one prepared PBCA acetone solution while maintaining 50/50 or 75/25 (w/w) and % of polymer in acetone at 2 and 3% (w/w), respectively. A mesh composed of high molecular weight PBCA with nanoparticles of magnesium encapsulated in particles of poly(lactic acid) was prepared by electrospinning of one emulsion formed by the addition of PLA-Mg powder. The fiber diameter of the mesh produced using a polymer concentration of 2% was around 360 nm, and when 3% was used, the diameter increased to 1 μm.

Example 20

Mechanical properties of electro-spun mesh with respect to molecular weight and polydispersity index of PBCA

As shown in Table 3 below, electro-spun meshes could be obtained from PBCAs with various molecular weights and polydispersity index values. Surprisingly, as shown in Table 3, PBCAs having a polydispersity index ranging from 1 to 1.7, such as PBCA No. An electro-spun mesh could also be obtained from 1.

Even more surprisingly, electro-spun meshes from PBCA with a polydispersity index ranging from 1 to 1.7 exhibited improved mechanical properties. As shown in Table 3, PBCA No. 1 and PBCA No. 1 having the same molecular weight but different polydispersity indices. It is revealed in the data of 2. Specifically, PBCA No. 1 having a polydispersity index in the range of 1 to 1.7. The mesh made from 1 had the same M _n , but with a polydispersity index falling outside the range of 1 to 1.7 PBCA No. It exhibited excellent mechanical properties when compared to the mesh prepared from 2. PBCA No. 1 having a polydispersity index ranging from 1 to 1.7. The mesh prepared from 1 had a fiber diameter of about 1 μm, a thickness of about 431 μm, a Young's modulus of about 1.4 MPa, and a tensile strength of about 0.07 MPa.

Example 21

PBCA mesh with fiber diameters ranging from 0.1 to 5 μm

As shown in FIGS. 14A-H , electro-spun meshes could be obtained from PBCA with fiber diameters ranging from 0.1 to 5 μm.

Claims (14)

(a) poly(alkyl cyanoacrylate) homopolymers and/or poly(alkyl cyanoacrylics) obtained by anionic polymerization of at least one alkyl cyanoacrylate (ACA) monomer and/or at least one ACA oligomer rate) copolymer, wherein the alkyl substituents of the ACA monomers and/or ACA oligomers may be optionally substituted, poly(alkyl cyanoacrylate) homopolymers and/or poly(alkyl cyanoacrylate) copolymers the ratio of the weight-average molecular weight (M _w ) to the number-average molecular weight (M _n ) of the coal is from 1 to 1.7;
(b) dissolving the poly(alkyl cyanoacrylate) homopolymer and/or poly(alkyl cyanoacrylate) copolymer of step (a) in a solvent;
(c) electrospinning the solution obtained in step (b) to obtain poly(alkyl cyanoacrylate)-based nano/microfibers on a collector; and
(d) taking out the poly(alkyl cyanoacrylate)-based nano/microfibers from the collector to obtain ready-to-use poly(alkyl cyanoacrylate)-based nano/microfibers.
A method for preparing ready-to-use poly(alkyl cyanoacrylate)-based nano/microfibers comprising: The poly(alkyl cyanoacrylate) copolymer of step (a) comprises (i) at least two different ACA monomer species, (ii) at least two different ACA monomer species. an alkyl cyanoacrylate oligomer, or (iii) at least one poly(alkyl cyanoacrylate) (PACA) polymer and at least one non-PACA polymer. 3. A compound according to claim 1 or 2, wherein at least one ACA monomer or at least two different ACA monomer species are n-butyl-2-cyanoacrylate (BCA), iso-butyl-2-cyanoacrylate (IBCA). ), n-pentyl-2-cyanoacrylate (PCA), iso-pentyl-2-cyanoacrylate (IPCA), n-hexyl-2-cyanoacrylate (HCA), iso-hexyl-2- cyanoacrylate (IHCA), cyclo-hexyl-2-cyanoacrylate (CHCA), n-heptyl-2-cyanoacrylate (HepCA), iso-heptyl-2-cyanoacrylate (IHepCA), selected from n-octyl-2-cyanoacrylate (OCA), iso-octyl-2-cyanoacrylate (IOCA), butyl-lactoyl-2-cyanoacrylate (BLCA) and mixtures thereof How to be. 3. The oligomer of claim 1 or 2, wherein the at least one ACA oligomer is an oligomer of n-butyl-2-cyanoacrylate (BCA), an oligomer of iso-butyl-2-cyanoacrylate (IBCA), n- Oligomers of pentyl-2-cyanoacrylate (PCA), oligomers of iso-pentyl-2-cyanoacrylate (IPCA), oligomers of n-hexyl-2-cyanoacrylate (HCA), iso-hexyl- oligomer of 2-cyanoacrylate (IHCA), oligomer of cyclo-hexyl-2-cyanoacrylate (CHCA), oligomer of n-heptyl-2-cyanoacrylate (HepCA), iso-heptyl-2- oligomer of cyanoacrylate (IHepCA), oligomer of n-octyl-2-cyanoacrylate (OCA), oligomer of iso-octyl-2-cyanoacrylate (IOCA), butyl-lactoyl-2-cya oligomers of noacrylate (BLCA) and mixtures thereof. 5. The solvent according to any one of claims 1 to 4, wherein the solvent of step (b) is acetone, ethanol, acetic acid, formic acid, ethyl acetate, dimethyl sulfoxide, dimethyl formamide, butyl acetate, isobutyl acetate and mixtures thereof. a method selected from any one of. 6 . The process according to claim 1 , wherein at least one non-PACA polymer is added to the solution obtained in step (b) prior to electrospinning. 7. The method of any one of claims 2-6, wherein the at least one non-PACA polymer is poly(lactic acid-co-glycolic acid) (PLGA), poly(ε-caprolactone) (PCL), poly(ethylene glycol) (PEG), poly(lactic acid) (PLA), chitosan (CH), hyaluronic acid (HA) and dextran (Dex). 8. The method according to any one of claims 1 to 7, wherein the ready-to-use nano/microfibers obtained in step (d) are not comprised in the support. The method according to any one of claims 1 to 8, wherein the ratio of the weight average molecular weight to the number average molecular weight is 1.1 to 1.5; or the ratio of the weight average molecular weight to the number average molecular weight is 1.2 to 1.4. A ready-to-use nano/microfiber obtainable by the method according to claim 1 . The ready-to-use nano/microfiber of claim 10 , wherein the fiber average diameter is between 100 nm and 5 micrometers. 11. The method of claim 10, wherein the therapeutic agent, antibiotic, antiviral agent, chemotherapeutic agent, analgesic and analgesic combination, anti-inflammatory agent, vitamin, sedative, radiopharmaceutical, metal particle, metal alloy particle, metal oxide particle, hydroxyapatite, sodium alginate, dye , cells, proteins, peptides, nucleic acids, nucleic acid analogs, nucleotides or oligonucleotides, peptide nucleic acids, aptamers, antibodies, or fragments or portions thereof, antigens or epitopes, hormones, hormone antagonists, growth factors or recombinant growth factors, and fragments thereof and ready-to-use nano/microfibers immobilized with or encapsulated therein with at least one of variants, cell adhesion mediators, cytokines, enzymes or mixtures thereof. Use of the ready-to-use nano/microfibers according to claim 10 in dental applications, for wound dressing/healing, tissue regeneration/engineering, organ protection, implant coating or controlled drug delivery. A nanofiber mesh comprising the ready-to-use nano/microfibers according to any one of claims 10 to 12, wherein the surface of the nanofiber mesh is formed by encapsulation and/or fixation of therapeutic agents and/or biological materials. A nanofiber mesh that is modified.

Images